Method for effectively protecting liguiniferous tissue of aquilaria tree from rotting and knotting

Abstract

This invention discloses an effective method for protecting the phloem tissue of agarwood trees from decay and resin formation, comprising the following steps: Selection of agarwood trees: Select agarwood trees over 3 years old with a diameter at breast height (DBH) of over 10 cm; Drill blind holes towards the tree's axis using a non-threaded drill bit, with a hole diameter of 5-8 mm and a depth of 4-5 cm; Arrange the holes in a stepped pattern from bottom to top, with a horizontal distance of 3-4 cm and a vertical distance of 1-2 cm between each hole and the next hole in the same row, and a vertical distance of 5-6 cm between adjacent rows; Introduce the resin-forming agent into the holes, and then apply a protective solution to the hole openings. This method effectively protects the phloem tissue of agarwood trees, achieving resin formation without decay. After resin formation, the heartwood accumulates yellowish-brown or reddish-brown resin, exhibiting a distinct agarwood layer structure, and the agarwood oil contains higher levels of effective components such as agaritol and agarwood aldehyde.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, specifically to a method for inducing resin formation in agarwood trees, and particularly to a method for effectively protecting the phloem tissue of agarwood trees from decay and resin formation. Background Technology

[0002] Agarwood is the resinous wood of plants in the Thymelaeaceae family, such as Aquilaria sinensis (Lour.) Gilg. Plants that can produce agarwood are collectively called agarwood trees. Due to the high value of agarwood, wild agarwood trees have been over-logged, leading to a growing scarcity of agarwood plant resources. To protect wild agarwood plants and develop sustainable agarwood production methods, various artificial cultivation techniques for agarwood trees and artificial resin formation have emerged.

[0003] When agarwood trees are struck by lightning, cut, or damaged by insects, their natural defense mechanisms trigger the secretion of resin, which over time forms high-quality agarwood. Utilizing this resin formation principle, early methods such as cutting, burning, nailing, and drilling were commonly used to treat agarwood trees, stimulating their defense mechanisms to promote resin production. However, these techniques are no longer suitable for modern technological advancements due to low yields, inconsistent quality, complex procedures, and negative impacts on tree health.

[0004] In recent years, the widely accepted method for agarwood formation is the drilling method, which only requires drilling holes in the tree trunk and then treating it with an agarwood-forming agent to quickly produce resin. This method effectively promotes resin formation; however, the traditional drilling method is destructive, damaging the phloem tissue. The phloem tissue, composed of sieve tubes and companion cells, sieve molecules, phloem fibers, and phloem parenchyma cells, is located between the bark and cambium. It can be divided into primary phloem and secondary phloem, and its main function is to transport nutrients. Damage to the phloem tissue affects nutrient transport in the agarwood tree, hindering its healthy growth. A good agarwood formation process involves preserving the phloem tissue or allowing it to heal rapidly, while the xylem tissue is inhibited or continuously damaged to promote resin formation and accumulation, resulting in high-quality, high-yield agarwood heartwood. Currently known drilling methods have not achieved these effects; therefore, this invention proposes an effective method for preventing the decay of the agarwood tree's phloem tissue during agarwood formation, which is of great significance. Summary of the Invention

[0005] In view of this, the present invention provides an effective method for protecting the phloem tissue of agarwood trees from decay and resin formation.

[0006] To achieve the above objectives, the present invention provides the following technical solution:

[0007] An effective method for protecting the phloem tissue of agarwood trees from decay and resin formation includes the following steps:

[0008] S1. Selection of Agarwood Trees: Select Agarwood trees that are 3 years or older and have a diameter at breast height of 10cm or more;

[0009] S2. Drilling: Drill blind holes in the direction of the tree axis using a threadless drill bit. The hole diameter is 5-8mm and the depth of the blind hole is 4-5cm. The holes in each row are arranged in a stepped manner from bottom to top. The horizontal distance between each hole and the next hole in the same row is 3-4cm, the vertical distance is 1-2cm, and the vertical distance between two adjacent rows is 5-6cm.

[0010] S3. Resin Formation: Introduce the resin-forming agent into the hole, and then apply a protective solution to the hole opening; the resin-forming agent is only used once in the initial stage of resin formation;

[0011] The fragrance-forming agent includes the following components: trans-cinnamic acid, potassium humate, anthocyanins, amino oligosaccharides, Rhodopseudomonas palustris bacterial solution or its fermentation supernatant, Beauveria bassiana bacterial solution, Paecilomyces lilacinus bacterial solution, glycine, manganese carbonate, ammonium molybdate, malic acid, and water.

[0012] Preferably, the fragrance-forming agent comprises, by weight percentage: 0.4%-0.8% trans-cinnamic acid, 2%-5% potassium humate, 1%-5% anthocyanins, 5%-10% amino oligosaccharides, 1.2%-1.7% *Rhodopseudomonas palustris* bacterial suspension and / or its fermentation supernatant, 1.5%-2.2% *Beauveria bassiana* bacterial suspension, 1.0%-1.6% *Paecilomyces lilacinus* bacterial suspension, 1%-3% glycine, 0.1%-0.5% manganese carbonate, 0.003%-0.008% ammonium molybdate, 0.03%-0.05% malic acid, and water.

[0013] Preferably, the fragrance-forming agent comprises, by weight percentage: 0.6% trans-cinnamic acid, 2% potassium humate, 3% anthocyanins, 8% amino oligosaccharides, 1.7% *Rhodopseudomonas palustris* fermentation supernatant, 1.5% *Beauveria bassiana* culture, 1.6% *Paecilomyces lilacinus* culture, 1% glycine, 0.1% manganese carbonate, 0.005% ammonium molybdate, and 0.03% malic acid and water.

[0014] Preferably, the method for preparing the fermentation supernatant of Rhodopseudomonas palustris includes the following steps:

[0015] Rhodopseudomonas palustris was inoculated into liquid culture medium and anaerobic fermented for 5-7 days at a temperature of 30℃~35℃ and a light intensity of 3000Lx~4000Lx. Then, the supernatant was collected by centrifugation.

[0016] Preferably, the liquid culture medium is formulated as follows: 6%-12% rosehip seed oil (CAS: 84603-93-0), 1%-3% propionic acid, 0.5%-0.7% yeast extract, 0.1%-0.5% chitosan, 0.05%-0.07% ammonium sulfate, 0.1%-1% disodium EDTA, 0.05%-0.1% ethanol, 0.05%-0.09% dipotassium hydrogen phosphate, 0-0.02% tocopherol, 0.02%-0.05% sodium chloride, and water.

[0017] Preferably, the liquid culture medium is formulated as follows: 8% rosehip oil, 2% propionic acid, 0.5% yeast extract, 0.5% chitosan, 0.05% ammonium sulfate, 0.8% disodium EDTA, 0.05% ethanol, 0.07% dipotassium hydrogen phosphate, 0.02% tocopherol, 0.02% sodium chloride, and water.

[0018] Compared with the prior art, the beneficial effects of the present invention are:

[0019] This invention utilizes a threadless drill bit with a small hole diameter to minimize damage to the phloem tissue of the agarwood tree during drilling. Furthermore, the improved resin-forming agent induces rapid resin formation in the agarwood tree and stimulates the phloem tissue's healing system. This allows the phloem tissue to heal quickly after the resin-forming agent is absorbed by the tree, ensuring that the agarwood tree continues to grow rapidly while simultaneously producing resin.

[0020] This invention minimizes damage to the tree without affecting resin formation, resulting in good economic benefits for growers. The heartwood after resin formation accumulates yellowish-brown or reddish-brown resin, exhibiting a distinct agarwood layer structure, and contains higher levels of effective components such as linalool and agarwood aldehyde in the agarwood oil.

[0021] The present invention uses a small amount of resin-forming agent and only requires one application of the resin-forming agent in the early stage of resin formation, without the need for secondary supplementation to induce resin formation. The method is simple and efficient. Attached Figure Description

[0022] Figures 1-3 Example 2 of this invention: Effect diagram of resin formation. Detailed Implementation

[0023] To enable those skilled in the art to better understand the technical content of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0024] Rhodopseudomonas palustris (Warner Bio), Pseudomonas stearothermia (Warner Bio), Beauveria bassiana (Zhongxiang Bio), Paecilomyces lilacinus (Zhongxiang Bio).

[0025] Example 1: An effective method for protecting the phloem tissue of agarwood trees from decay and resin formation.

[0026] S1. Selection of Agarwood Trees: Select Agarwood trees that are 3 years or older and have a diameter at breast height (diameter at a height of 1.3 meters above the ground) of 10 cm or more;

[0027] S2. Drilling: Start drilling from a height of 1.2 meters or above. Each row of holes is arranged in a stepped manner from bottom to top. The horizontal distance between each hole and the next hole (in the same row) is 3-4 cm, and the vertical distance is 1-2 cm. The vertical distance between two adjacent rows is 5-6 cm. After selecting the hole position, use a non-threaded drill bit to drill blind holes in the direction of the tree axis. The hole diameter is 5-8 mm, and the depth of the blind hole is 4-5 cm.

[0028] S3. Resin Formation: Introduce the resin-forming agent into the holes, using 10 mL per hole, and then apply a conditioning solution to the hole opening. The resin-forming agent should only be used once during the initial stage of resin formation.

[0029] The fragrance-forming agent includes the following components: 0.6% trans-cinnamic acid, 2% potassium humate, 3% anthocyanins, 8% amino oligosaccharides, and 1.7% *Rhodopseudomonas palustris* bacterial suspension (10... 8 CFU / mL), 1.5% Beauveria bassiana bacterial suspension (10 8 cfu / mL), 1.6% Paecilomyces lilacinus bacterial solution (10 8 The formula consists of cfu / mL, 1% glycine, 0.1% manganese carbonate, 0.005% ammonium molybdate, 0.03% malic acid, and water.

[0030] The care solution comprises the following ingredients: 3% methyl jasmonate, 17% calcium carbonate, 0.1% hydroxymethyl cellulose, and water.

[0031] Example 2: An effective method for protecting the phloem tissue of agarwood trees from decay and resin formation.

[0032] S1. Selection of Agarwood Trees: Select Agarwood trees that are 3 years or older and have a diameter at breast height (diameter at a height of 1.3 meters above the ground) of 10 cm or more;

[0033] S2. Drilling: Start drilling from a height of 1.2 meters or above. Each row of holes is arranged in a stepped manner from bottom to top. The horizontal distance between each hole and the next hole (in the same row) is 3-4 cm, and the vertical distance is 1-2 cm. The vertical distance between two adjacent rows is 5-6 cm. After selecting the hole position, use a non-threaded drill bit to drill blind holes in the direction of the tree axis. The hole diameter is 5-8 mm, and the depth of the blind hole is 4-5 cm.

[0034] S3. Resin Formation: Introduce the resin-forming agent into the holes, using 10 mL per hole, and then apply a conditioning solution to the hole opening. The resin-forming agent should only be used once during the initial stage of resin formation.

[0035] The fragrance-forming agent comprises the following components: 0.4%-0.8% trans-cinnamic acid (preferably 0.6% in this example), 2%-5% potassium humate (preferably 2% in this example), 1%-5% anthocyanins (preferably 3% in this example), 5%-10% amino oligosaccharides (preferably 8% in this example), 1.2%-1.7% *Rhodopseudomonas palustris* fermentation supernatant (preferably 1.7% in this example), and 1.5%-2.2% *Beauveria bassiana* bacterial suspension. 8 CFU / mL (preferably 1.5% in this example), 1.0%-1.6% Paecilomyces lilacinus bacterial solution 10 8 CFU / mL (preferably 1.6% in this example), 1%-3% glycine (preferably 1% in this example), 0.1%-0.5% manganese carbonate (preferably 0.1% in this example), 0.003%-0.008% ammonium molybdate (preferably 0.005% in this example), 0.03%-0.05% malic acid (preferably 0.03% in this example), and water.

[0036] The care solution comprises the following ingredients: 3% methyl jasmonate, 17% calcium carbonate, 0.1% hydroxymethyl cellulose, and water.

[0037] Preparation method of fermentation supernatant of Rhodopseudomonas palustris: Rhodopseudomonas palustris was prepared at a concentration of 1×10⁻⁶. 7 Inoculate the liquid culture medium with an inoculum of cfu / mL, and anaerobic ferment for 5 days at a temperature of 30℃~35℃ and a light condition of 3000Lx~4000Lx. Then centrifuge at 6000rpm for 10min, collect the supernatant and store it for later use.

[0038] The liquid culture medium is formulated as follows: 6%-12% rosehip oil (preferably 8% in this example), 1%-3% propionic acid (preferably 2% in this example), 0.5%-0.7% yeast extract (preferably 0.5% in this example), 0.1%-0.5% chitosan (preferably 0.5% in this example), 0.05%-0.07% ammonium sulfate (preferably 0.05% in this example), 0.1%-1% disodium ethylenediaminetetraacetate (preferably 0.8% in this example), 0.05%-0.1% ethanol (preferably 0.05% in this example), 0.05%-0.09% dipotassium hydrogen phosphate (preferably 0.07% in this example), 0-0.02% tocopherol (preferably 0.02% in this example), 0.02%-0.05% sodium chloride (preferably 0.02% in this example), and water.

[0039] Example 3: An effective method for protecting the phloem tissue of agarwood trees from decay and resin formation.

[0040] S1. Selection of Agarwood Trees: Select Agarwood trees that are 3 years or older and have a diameter at breast height (diameter at a height of 1.3 meters above the ground) of 10 cm or more;

[0041] S2. Drilling: Start drilling from a height of 1.2 meters or above. Each row of holes is arranged in a stepped manner from bottom to top. The horizontal distance between each hole and the next hole (in the same row) is 3-4 cm, and the vertical distance is 1-2 cm. The vertical distance between two adjacent rows is 5-6 cm. After selecting the hole position, use a non-threaded drill bit to drill blind holes in the direction of the tree axis. The hole diameter is 5-8 mm, and the depth of the blind hole is 4-5 cm.

[0042] S3. Resin Formation: Introduce the resin-forming agent into the holes, using 10 mL per hole, and then apply a conditioning solution to the hole opening. The resin-forming agent should only be used once during the initial stage of resin formation.

[0043] The fragrance-forming agent includes the following components: 0.6% trans-cinnamic acid, 2% potassium humate, 3% anthocyanins, 8% amino oligosaccharides, and 1.7% *Pseudomonas stearothermii* bacterial suspension (10... 8 CFU / mL), 1.5% Beauveria bassiana bacterial suspension (10 8 cfu / mL), 1.6% Paecilomyces lilacinus bacterial solution (10 8 The formula consists of cfu / mL, 1% glycine, 0.1% manganese carbonate, 0.005% ammonium molybdate, 0.03% malic acid, and water.

[0044] The care solution comprises the following ingredients: 3% methyl jasmonate, 17% calcium carbonate, 0.1% hydroxymethyl cellulose, and water.

[0045] Example 4: An effective method for protecting the phloem tissue of agarwood trees from decay and resin formation.

[0046] S1. Selection of Agarwood Trees: Select Agarwood trees that are 3 years or older and have a diameter at breast height (diameter at a height of 1.3 meters above the ground) of 10 cm or more;

[0047] S2. Drilling: Start drilling from a height of 1.2 meters or above. Each row of holes is arranged in a stepped manner from bottom to top. The horizontal distance between each hole and the next hole (in the same row) is 3-4 cm, and the vertical distance is 1-2 cm. The vertical distance between two adjacent rows is 5-6 cm. After selecting the hole position, use a non-threaded drill bit to drill blind holes in the direction of the tree axis. The hole diameter is 5-8 mm, and the depth of the blind hole is 4-5 cm.

[0048] S3. Resin Formation: Introduce the resin-forming agent into the holes, using 10 mL per hole, and then apply a conditioning solution to the hole opening. The resin-forming agent should only be used once during the initial stage of resin formation.

[0049] The fragrance-forming agent includes the following components: 0.6% trans-cinnamic acid, 2% potassium humate, 3% anthocyanins, 8% amino oligosaccharides, 1.7% *Rhodopseudomonas palustris* fermentation supernatant, and 1.5% *Beauveria bassiana* bacterial suspension (10... 8 cfu / mL), 1.6% Paecilomyces lilacinus bacterial solution (10 8The formula consists of cfu / mL, 1% glycine, 0.1% manganese carbonate, 0.005% ammonium molybdate, 0.03% malic acid, and water.

[0050] The care solution comprises the following ingredients: 3% methyl jasmonate, 17% calcium carbonate, 0.1% hydroxymethyl cellulose, and water.

[0051] Preparation method of fermentation supernatant of Rhodopseudomonas palustris: Rhodopseudomonas palustris was prepared at a concentration of 1×10⁻⁶. 7 Inoculate the liquid culture medium with an inoculum of cfu / mL, and anaerobic ferment for 5 days at a temperature of 30℃~35℃ and a light condition of 3000Lx~4000Lx. Then centrifuge at 6000rpm for 10min, collect the supernatant and store it for later use.

[0052] The liquid culture medium formula is as follows: 10% propionic acid, 0.5% yeast extract, 0.5% chitosan, 0.05% ammonium sulfate, 0.8% disodium EDTA, 0.05% ethanol, 0.07% dipotassium hydrogen phosphate, 0.02% tocopherol, 0.02% sodium chloride, and water.

[0053] Example 5: An effective method for protecting the phloem tissue of agarwood trees from decay and resin formation.

[0054] S1. Selection of Agarwood Trees: Select Agarwood trees that are 3 years or older and have a diameter at breast height (diameter at a height of 1.3 meters above the ground) of 10 cm or more;

[0055] S2. Drilling: Start drilling from a height of 1.2 meters or above. Each row of holes is arranged in a stepped manner from bottom to top. The horizontal distance between each hole and the next hole (in the same row) is 3-4 cm, and the vertical distance is 1-2 cm. The vertical distance between two adjacent rows is 5-6 cm. After selecting the hole position, use a non-threaded drill bit to drill blind holes in the direction of the tree axis. The hole diameter is 5-8 mm, and the depth of the blind hole is 4-5 cm.

[0056] S3. Resin Formation: Introduce the resin-forming agent into the holes, using 10 mL per hole, and then apply a conditioning solution to the hole opening. The resin-forming agent should only be used once during the initial stage of resin formation.

[0057] The fragrance-forming agent comprises the following components: 0.4%-0.8% trans-cinnamic acid (preferably 0.6% in this example), 2%-5% potassium humate (preferably 2% in this example), 1%-5% anthocyanins (preferably 3% in this example), 5%-10% amino oligosaccharides (preferably 8% in this example), 1.2%-1.7% *Rhodopseudomonas palustris* fermentation supernatant (preferably 1.7% in this example), and 1.0%-1.6% *Paecilomyces lilacinus* bacterial solution. 8CFU / mL (preferably 1.6% in this example), 1%-3% glycine (preferably 1% in this example), 0.1%-0.5% manganese carbonate (preferably 0.1% in this example), 0.003%-0.008% ammonium molybdate (preferably 0.005% in this example), 0.03%-0.05% malic acid (preferably 0.03% in this example), and water.

[0058] The care solution comprises the following ingredients: 3% methyl jasmonate, 17% calcium carbonate, 0.1% hydroxymethyl cellulose, and water.

[0059] Preparation method of fermentation supernatant of Rhodopseudomonas palustris: Rhodopseudomonas palustris was prepared at a concentration of 1×10⁻⁶. 7 Inoculate the liquid culture medium with an inoculum of cfu / mL, and anaerobic ferment for 5 days at a temperature of 30℃~35℃ and a light condition of 3000Lx~4000Lx. Then centrifuge at 6000rpm for 10min, collect the supernatant and store it for later use.

[0060] The liquid culture medium is formulated as follows: 6%-12% rosehip oil (preferably 8% in this example), 1%-3% propionic acid (preferably 2% in this example), 0.5%-0.7% yeast extract (preferably 0.5% in this example), 0.1%-0.5% chitosan (preferably 0.5% in this example), 0.05%-0.07% ammonium sulfate (preferably 0.05% in this example), 0.1%-1% disodium ethylenediaminetetraacetate (preferably 0.8% in this example), 0.05%-0.1% ethanol (preferably 0.05% in this example), 0.05%-0.09% dipotassium hydrogen phosphate (preferably 0.07% in this example), 0-0.02% tocopherol (preferably 0.02% in this example), 0.02%-0.05% sodium chloride (preferably 0.02% in this example), and water.

[0061] Experimental Example: Quality Testing of Agarwood

[0062] Agarwood was collected 3 months after injection of the resin-forming agent. The content of extractives, linalool content, and characteristic peaks were determined according to the "Extractives," "Content Determination," and "Characteristic Spectra" sections of the Chinese Pharmacopoeia 2020, Part I, Agarwood, with wild agarwood as a control. Agarwood oil was extracted, and the content of agarwood aldehyde in the oil was determined according to the reported GC-MS method (see Table 1).

[0063] Gas chromatography conditions:

[0064] The chromatographic column was a Zebron ZB-5MSI 5% Phenyl-95% Dimethylpolysiloxane (30m×0.25mm×0.25μm) flexible quartz capillary column. The temperature program was: 50℃ for 2 min, then increased to 320℃ at 5℃ / min and held for 3 min. High-purity helium was used as the carrier gas. The column inlet pressure was 7.6 psi, and the carrier gas flow rate was 1.0 mL / min. The injection method was split injection with a split ratio of 40:1 and a solvent delay time of 4 min.

[0065] The EI source is an ion source with an ion source temperature of 230℃, a quadrupole temperature of 150℃, an electron energy of 70eV, an emission current of 34.6μA, a multiplier voltage of 1108V, an interface temperature of 280℃, and a mass range of 20–450 amu.

[0066] Mass spectra were obtained after mass spectrometry scanning. The types of compounds were identified by searching the mass spectrometry database system, and the relative content of the compounds was determined by peak area normalization.

[0067] Table 1 n=10

[0068] aldehyde content of white wood Agaric tetraol content Example 1 6.7％ 0.28％ Example 2 8.1％ 0.44％ Example 3 4.4％ 0.37％ Example 4 7.3％ 0.33％ Example 5 7.0％ 0.40％ Wild Agarwood 2.9％ 0.24％

[0069] The test results showed that all agarwood samples from each embodiment had six characteristic peaks corresponding to the control chromatographic peaks. The extract content of all agarwood samples from each embodiment reached more than 25%. The content of agarotetraol was greater than 0.1%, meeting the quality requirements. Among them, Example 2 had the highest agarotetraol content, significantly higher than the other groups (P < 0.05).

[0070] Compared to Example 4, Example 2, which uses rosehip oil as the dominant carbon source for fermentation, has a significantly higher content of the active ingredient agaric aldehyde in the volatile oil after resin formation compared to other examples.

[0071] Compared to Example 2, Example 5 did not add Beauveria bassiana liquid, and the contents of agaric tetraol and agaric aldehyde were reduced, indicating that Beauveria bassiana can also induce agarwood formation in Aquilaria sinensis.

[0072] The above description is only a part of the embodiments of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention shall fall within the protection scope of the present invention.

Claims

1. A method for effectively protecting the phloem tissue of agarwood trees from decay and resin formation, characterized in that, Includes the following steps: S1. Selection of Agarwood Trees: Select Agarwood trees that are 3 years or older and have a diameter at breast height of 10cm or more; S2. Drilling: Drill blind holes in the direction of the tree axis using a threadless drill bit. The hole diameter is 5-8mm and the depth of the blind hole is 4-5cm. The holes in each row are arranged in a stepped manner from bottom to top. The horizontal distance between each hole and the next hole in the same row is 3-4cm and the vertical distance is 1-2cm. The vertical distance between two adjacent rows is 5-6cm. S3. Resin Formation: Introduce the resin-forming agent into the hole, and then apply a protective solution to the hole opening; the resin-forming agent is only used once in the initial stage of resin formation; By weight percentage, the fragrance-forming agent comprises the following components: 0.6% trans-cinnamic acid, 2% potassium humate, 3% anthocyanins, 8% amino oligosaccharides, 1.7% *Rhodopseudomonas palustris* fermentation supernatant, 1.5% *Beauveria bassiana* culture, 1.6% *Paecilomyces lilacinus* culture, 1% glycine, 0.1% manganese carbonate, 0.005% ammonium molybdate, and 0.03% malic acid and water.

2. The method for effectively protecting the phloem tissue of agarwood trees from decay and resin formation according to claim 1, characterized in that, The method for preparing fermentation supernatant of Rhodopseudomonas palustris includes the following steps: Rhodopseudomonas palustris was inoculated into liquid culture medium and anaerobic fermented for 5-7 days at a temperature of 30℃~35℃ and a light intensity of 3000Lx~4000Lx. Then, the supernatant was collected by centrifugation.

3. The method for effectively protecting the phloem tissue of agarwood trees from decay and resin formation according to claim 2, characterized in that, The liquid culture medium is formulated as follows: 6%-12% rosehip oil, 1%-3% propionic acid, 0.5%-0.7% yeast extract, 0.1%-0.5% chitosan, 0.05%-0.07% ammonium sulfate, 0.1%-1% disodium EDTA, 0.05%-0.1% ethanol, 0.05%-0.09% dipotassium hydrogen phosphate, 0-0.02% tocopherol, 0.02%-0.05% sodium chloride, and water.

4. The method for effectively protecting the phloem tissue of agarwood trees from decay and resin formation according to claim 2, characterized in that, The liquid culture medium is formulated as follows: 8% rosehip oil, 2% propionic acid, 0.5% yeast extract, 0.5% chitosan, 0.05% ammonium sulfate, 0.8% disodium EDTA, 0.05% ethanol, 0.07% dipotassium hydrogen phosphate, 0.02% tocopherol, 0.02% sodium chloride, and water.

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