A method for promoting agarwood production

Abstract

The present invention discloses a method for promoting agarwood production in agarwood trees, comprising the following steps: artificially producing agarwood by inoculating the agarwood mouth with a single strain of anomalous Wickham yeast. The anomalous Wickham yeast was purchased from the Shanghai Collection of Microorganisms, strain number SHBCC D53500. This method requires only a single inoculation of the anomalous Wickham yeast, resulting in agarwood production within a short timeframe. The agarwood matures quickly, and its aroma resembles that of wild agarwood.

Description

Technical Field

[0001] The present invention relates to the field of microbial applications, and in particular to a method for promoting agarwood production in agarwood trees. Background Art

[0002] Agarwood is a solid product produced by a special type of agarwood tree, a mixture of oil (resin) and wood. The wood of this type of agarwood tree itself has no distinctive fragrance and is relatively soft. Several trees in the genus Aquilaria (Thymelaeaceae), such as the white agarwood tree and the agarwood tree, can produce agarwood. However, not every agarwood tree produces agarwood. During its growth, agarwood trees can sustain wounds from animal bites, external trauma, human damage, and erosion by snakes, insects, and ants. However, these wounds do not heal quickly. To prevent moisture evaporation and microbial infection, the agarwood tree spontaneously secretes resin to protect the wound. Under the action of the agarwood tree's oils and bacteria, the wound gradually forms a paste-like mass. As the wound heals, new wood covers it, creating a resinous layer. Gradually, the wound becomes agarwood.

[0003] Agarwood trees have relatively high requirements for their growing environment, and whether they can produce agarwood depends on chance. Moreover, it takes dozens or even hundreds of years for agarwood to form under natural conditions and long-term sedimentation.

[0004] In recent years, the industry has conducted a lot of research on artificially planted agarwood trees and artificial agarwood, and has achieved great success, shortening the formation time of agarwood and increasing the yield of agarwood. However, there are still problems such as long agarwood formation time, long maturity time, low yield, and the fragrance is quite different from wild agarwood. Summary of the Invention

[0005] In order to overcome the above-mentioned shortcomings and deficiencies of the prior art, the object of the present invention is to provide a method for promoting the agarwood production of agarwood trees, which only requires a one-time inoculation of abnormal Wickham yeast, can produce agarwood in a short time, the agarwood matures in a short time, and the fragrance is close to that of wild agarwood.

[0006] The purpose of the present invention is achieved through the following technical solutions:

[0007] A method for promoting agarwood production, comprising the following steps:

[0008] The agarwood mouth of the agarwood tree is inoculated with abnormal Wickham yeast at one time to artificially form agarwood.

[0009] Preferably, the abnormal Wickham yeast was purchased from Shanghai Collection of Microorganisms, with the strain number SHBCCD53500.

[0010] Preferably, the artificial incense takes less than 15 days to form and less than 3 years to mature.

[0011] Preferably, the incense mouth is formed by drilling or sawing.

[0012] Preferably, the agarwood tree is a white agarwood tree, and the extract of the obtained mature agarwood contains white agarwood aldehyde, α-santalol, α-costus aldehyde, and isowhite agarwood alcohol.

[0013] Preferably, the artificial incense formation specifically comprises the following steps:

[0014] (1) Bacterial culture:

[0015] The abnormal Wickham yeast was cultured in a culture medium until the logarithmic growth period at a culture temperature of 25 to 28° C. to obtain an abnormal Wickham yeast culture liquid;

[0016] (2) Open the mouth;

[0017] (3) Inoculation: Take the abnormal Wickham yeast liquid in step (1) and inoculate it into the fragrant gate.

[0018] Preferably, in the step of opening the incense mouth in step (2), the agarwood tree is a white agarwood tree, and the incense mouth of each white agarwood tree is opened 1.5 to 2 meters from the ground by sawing; the distance between the incense mouths on the same white agarwood tree is 30 to 40 cm, and the incense mouth width is 1 to 4 cm.

[0019] Preferably, in the step (2) of opening the incense mouth, the agarwood tree is a Kynam tree, and holes are drilled on the trunk or side branches. The incense mouths are small circular holes with a depth of 1 / 2 to 3 / 5 of the trunk diameter; the distance between adjacent incense mouths is 9 to 11 cm; when drilling holes on the trunk, the diameter of the holes is 0.6 to 1.0 cm; when drilling holes on the side branches, the diameter of the holes is 0.3 to 0.5 cm.

[0020] Preferably, in step (3) the inoculation step, the site is sealed with a plastic film after inoculation, and is cared for by spraying or misting for moisturizing every two months.

[0021] Preferably, the agarwood tree is Aquilaria sinensis or Aquilaria sinensis.

[0022] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0023] The invention inoculates abnormal Wickham yeast at the agarwood mouth of the agarwood tree at one time, so that agarwood can be produced in 10 to 15 days and the maturity period is about 2 years. The agarwood has high yield and high quality, and the fragrance is close to that of wild agarwood. The yield of agarwood is greatly increased, the production time is shortened, and the operation is simple. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is a diagram showing the aroma formation effect of the experimental group samples of Example 1 of the present invention 13 days after being inoculated with abnormal Wickham yeast.

[0025] Figure 2 This is a diagram showing the fragrance formation effect of the control group sample of Example 1 of the present invention one and a half years after the fragrance mouth was opened.

[0026] Figure 3 This is a diagram showing the effect of one year of fragrance formation by drilling holes on small side branches and inoculating the abnormal Wickham yeast of this example in Example 2 of the present invention.

[0027] Figure 4 This is a diagram showing the effect of drilling holes in tree trunks for fragrance without inoculating fungi for one year according to Example 2 of the present invention. DETAILED DESCRIPTION

[0028] The present invention will be further described in detail below with reference to the examples, but the embodiments of the present invention are not limited thereto.

[0029] Example 1

[0030] Among the Aquilaria sinensis trees planted at the Lingao base in Hainan, 10 Aquilaria sinensis trees with an age of 8 years, a tree diameter of 12 cm, good growth and similar growth status were selected and divided into two groups. The experimental group (5 trees, numbered 1 to 5) was inoculated with the abnormal Wickham yeast of the present invention (purchased from Shanghai Collection of Microorganisms, strain number SHBCC D53500) for artificial fragrance formation.

[0031] The specific implementation process of the experimental group is as follows:

[0032] (11) Bacterial culture:

[0033] The abnormal Wickham yeast (SHBCC D53500) was cultured in a culture medium until the logarithmic growth cycle at a culture temperature of 25-28° C. to obtain an abnormal Wickham yeast culture liquid;

[0034] The components of the culture medium include malt extract and agar, and the preparation method is as follows:

[0035] (12) Preparation of malt wort: Take several barley malts, crush them, add 3 times the weight of water to the malts, stir evenly, saccharify in a water bath at 58°C for 3 hours, filter until the iodine does not turn blue, add egg white, boil and filter to obtain malt wort.

[0036] (13) Preparation of 50Be malt extract agar: Dilute the malt extract with water to 50Be, add 15g agar, and sterilize at 121℃ for 20 minutes. The pH is natural.

[0037] (2) Open the fragrance mouth:

[0038] In the experimental group, the incense mouth of each individual Aquilaria sinensis tree was opened 2m from the ground; the size and shape of the incense mouth of each tree were basically the same: the distance between the incense mouths on the same Aquilaria sinensis tree was about 30cm, and the incense mouth was 3cm wide.

[0039] (3) Vaccination:

[0040] Take 5 ml of the abnormal Wickham yeast liquid in step (1), inoculate it into the incense gate, and seal it with plastic film.

[0041] (4) Nursing:

[0042] Spray or mist moisturizer every two months.

[0043] The implementation process of the control group is as follows:

[0044] The steps for opening the fragrance mouth were the same as those in the experimental group.

[0045] The experimental group and the control group were observed. The time when the white agarwood trees numbered 1 to 5 in the experimental group began to produce agarwood (with obvious formation of linear agarwood substances) was 13, 12, 13, and 14 days respectively; while in the control group, none of them produced agarwood after one and a half years.

[0046] Figure 1 This is a diagram of the incense formation effect of the experimental group samples of this embodiment 13 days after inoculation with abnormal Wickham yeast. Linear agarwood material (incense thread) is clearly formed at the incense mouth.

[0047] Figure 2 This is a picture of the incense effect of the control group sample of this embodiment one and a half years after it was opened. There is almost no incense line visible at the incense mouth.

[0048] The experimental group samples (grown for two years) of this embodiment were tested, and the results obtained were as follows:

[0049] Extract content: 52.41%; Detection method: Pharmacopoeia of the People's Republic of China (2020).

[0050] Thin layer chromatography identification: The color spots are the same as those of the agarwood reference substance; detection method: Pharmacopoeia of the People's Republic of China (2020).

[0051] The extract of this embodiment was subjected to gas chromatography-mass spectrometry analysis, and the results showed that the total content of chromones in the extract of this embodiment was 45.73%, of which the total content of 2-(2-phenylethyl)chromone compounds was 5.84%; it contained agarwood aldehyde, α-santalol, α-costus aldehyde, and isoagarwood alcohol; the content of the characteristic component agarwood aldehyde was 4.37%; and the fragrance was similar to that of wild white agarwood.

[0052] Example 2

[0053] Three-year-old agarwood trees were artificially wounded by drilling, and the anomalous Wickham yeast of the present invention (purchased from Shanghai Collection of Microorganisms, strain number SHBCC D53500) was inoculated to artificially form agarwood.

[0054] The specific implementation process is as follows:

[0055] (11) Bacterial culture:

[0056] The abnormal Wickham yeast (SHBCC D53500) was cultured in a culture medium until the logarithmic growth cycle at a culture temperature of 25-28° C. to obtain an abnormal Wickham yeast culture liquid;

[0057] The components of the culture medium include malt extract and agar, and the preparation method is as follows:

[0058] (12) Preparation of malt wort: Take several barley malts, crush them, add 4 times the weight of water to the malts, stir evenly, saccharify in a 60℃ water bath for 3.5 hours, filter until the iodine is no longer blue, add egg white, boil and filter to obtain malt wort.

[0059] (13) Preparation of 50Be malt extract agar: Dilute the malt extract with water to 50Be, add 15g agar, and sterilize at 121℃ for 20 minutes. The pH is natural.

[0060] (2) Open the fragrance mouth:

[0061] Drill holes on the agarwood tree to create scent holes. The scent holes can be made at a distance of more than 1.5m from the trunk or on a side branch. The scent holes are small round holes with a depth of 1 / 2 the diameter of the trunk or side branch. The distance between adjacent scent holes is 10cm. When drilling on the trunk, the diameter of the holes is 0.6-1.0cm; when drilling on a side branch, the diameter of the holes is 0.3-0.5cm.

[0062] (3) Vaccination:

[0063] Take 5 ml of the abnormal Wickham yeast liquid in step (1), inoculate it into the incense gate, and seal it with plastic film.

[0064] (4) Nursing:

[0065] Spray or mist moisturizer every two months.

[0066] Figure 3 This is the effect of drilling holes on small side branches and inoculating the abnormal Wickham yeast of this embodiment for one year; while the holes drilled on the trunk were not inoculated with the fungus and no fragrance was produced in one year. Figure 4 .

[0067] The agarwood grown for two years in this embodiment was tested, and the results obtained were as follows:

[0068] Extract content: 38.94%; Detection method: Pharmacopoeia of the People's Republic of China (2020).

[0069] Thin layer chromatography identification: The color spots are the same as those of the agarwood reference substance; detection method: Pharmacopoeia of the People's Republic of China (2020).

[0070] Gas chromatography-mass spectrometry analysis of the extract of this example showed that the total content of chromones in the extract of this example was 82.30%, of which the total content of 2-(2-phenylethyl)chromone compounds was 37.06%; the content of (-)guaiac-1(10),11-dien-15-ol was 0.78%; the content of white agarwood aldehyde was 1.02%; and the content of α-santalol was 1.05%. Its fragrance is similar to that of wild agarwood.

[0071] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the 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. A method for promoting agarwood production, characterized in that: The following steps are involved: The agarwood tree is inoculated with abnormal Wickham yeast at one time to artificially form agarwood; The abnormal Wickham yeast was purchased from Shanghai Collection of Microorganisms, strain number SHBCC D53500; The artificial incense has a formation time of less than 15 days and a maturity time of less than 3 years; The artificial incense formation method specifically comprises the following steps: (1) Bacterial culture: The abnormal Wickham yeast was cultured in a culture medium until the logarithmic growth period at a culture temperature of 25-28°C to obtain an abnormal Wickham yeast culture liquid; (2) Open the fragrance mouth; (3) Inoculation: Take the abnormal Wickham yeast liquid in step (1) and inoculate it into the fragrant gate.

2. The method for promoting agarwood production according to claim 1, wherein: The incense mouth is formed by drilling or sawing.

3. The method for promoting agarwood production according to claim 1, wherein: In the step of opening the incense mouth in step (2), the agarwood tree is a white agarwood tree, and the incense mouth of each white agarwood tree is opened 1.5 to 2 meters from the ground by sawing; the distance between the incense mouths on the same white agarwood tree is 30 to 40 cm, and the incense mouth width is 1 to 4 cm.

4. The method for promoting agarwood production according to claim 1, wherein: In the step of opening the incense mouth in step (2), the agarwood tree is a Kynam agarwood tree, and a hole is drilled on the trunk or side branches. The incense mouth is a small circular hole with a depth of 1 / 2 to 3 / 5 of the trunk diameter; the distance between adjacent incense mouths is 9 to 11 cm; when drilling on the trunk, the diameter of the small hole is 0.6 to 1.0 cm; when drilling on the side branches, the diameter of the small hole is 0.3 to 0.5 cm.

5. The method for promoting agarwood production according to claim 1, wherein: In step (3) of the inoculation step, the inoculation is followed by wrapping and sealing with plastic film, and then performing care: spraying or misting for moisturizing every two months.

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