Method for planting white truffle in pinus massoniana

Abstract

The application provides a method for planting white mushroom on Smith pine stumps, and relates to the technical field of artificial planting of white mushroom. The method for planting white mushroom on Smith pine stumps comprises the following steps: sterilizing old stumps / new stumps after felling of Smith pine by using a blowtorch, pouring a water solution of extract of salt water of drunken fish grass, and inoculating liquid spawn of white mushroom after air-drying, wherein the liquid spawn of white mushroom is obtained by using a mixture of almond paste, extract of salt water of drunken fish grass and potato liquid medium as a culture medium. The application overcomes the shortcomings of the prior art, effectively promotes the growth of white mushroom on the stumps by sterilizing the stumps and using the nutrient agent, adjusts the culture medium components of the white mushroom, increases the yield, and improves the economic benefits.

Description

Technical Field

[0001] This invention relates to the field of artificial cultivation technology of white ginseng fungus, specifically to a method for cultivating white ginseng fungus by felling pine stumps. Background Technology

[0002] Simao pine is an evergreen tree belonging to the genus Pinus of the family Pinaceae. Because its trunk is straight and not twisted, and its growth cycle is relatively short, it is usually used for timber. In Simao pine forests, there are usually many new and old tree stumps after logging.

[0003] White ginseng fungus, also known as split-gill fungus, white flower, and eight-dan firewood, is a precious edible and medicinal fungus belonging to the kingdom Fungi, phylum Basidiomycota, and family Split-gillaceae. Its cap is fan-shaped or kidney-shaped, 1-5 cm in diameter, white to grayish-white in color, with numerous lobes. The flesh is thin yet firm, with a delicate and delicious flavor and rich nutritional value. As a wood-rotting fungus, wild white ginseng is generally found on the dead branches and fallen logs of broad-leaved trees (such as oak, sago palm, and chestnut) and coniferous trees. This results in limited wild white ginseng production, unable to meet market demand. Therefore, artificial cultivation of white ginseng is a major development direction at present. Currently, artificial cultivation mainly uses cellulose-rich agricultural byproducts such as cottonseed hulls, corn cobs, sugarcane bagasse, and sawdust as the main materials, supplemented with wheat bran, rice bran, and trace minerals as the culture medium. It requires indoor greenhouse cultivation. For the Simao pine forests in Yunnan, numerous new and old tree stumps can serve as natural culture media for white ginseng. However, direct inoculation results in low overall yield and low economic value. Therefore, how to utilize the residue after logging Simao pine for white ginseng cultivation to improve the economic benefits of cultivation is a major research direction at present. Summary of the Invention

[0004] To address the shortcomings of existing technologies, this invention provides a method for cultivating white ginseng fungus on tree stumps. By sterilizing the tree stumps, using nutrient agents, and adjusting the composition of the white ginseng fungus culture medium, the method effectively promotes the growth of white ginseng fungus on the tree stumps, increases yield, and improves economic benefits.

[0005] To achieve the above objectives, the present invention provides the following technical solution:

[0006] A method for cultivating white ginseng fungus on Pinus simonii stumps, the method comprising the following steps:

[0007] S1. Use new or old stumps of Pinus simonii after felling as planting material. Two months before the start of the rainy season, use a blowtorch to sterilize the surface of the new or old stumps to obtain sterilized stumps for later use.

[0008] S2. Select the fruiting bodies of white ginseng that grow naturally on dead trees of Pinus sylvestris in the Pinus sylvestris forest as the strain, and inoculate the strain into potato protein medium to cultivate mycelium;

[0009] S3. The mycelium is mixed with sterile water and ground to prepare a suspension. The suspension is then inoculated into a liquid culture medium with the following formula: 10-20 mL almond milk + 600-800 mg Buddleja officinalis salt extract + 1 L potato liquid culture medium. This yields a liquid culture.

[0010] S4. Prepare a nutrient solution of 200-400 mg / L of Buddleja officinalis salt extract. On a sunny day, thoroughly pour the sterilized nutrient solution onto the surface of the sterilized pile material once, and then let it air dry naturally to obtain the pre-treated pile material for later use.

[0011] S5. Pour the liquid inoculum onto the pretreated pile material to complete the inoculation. Harvest 3-4 months later.

[0012] Preferably, in step S1, after the blowtorch sterilizes the surface of the new or old stake, the vegetation around the new or old stake is also sterilized, with the treatment range being 20-50cm in radius of the new or old stake.

[0013] Preferably, the potato protein culture medium is formulated as follows: 200g potato + 20g glucose + 3g peptone + 2g potassium dihydrogen phosphate + 18g agar + 0.5g magnesium sulfate + 1L water.

[0014] Preferably, the formula of the potato liquid culture medium is 200g potato + 20g glucose + 3g peptone + 2g potassium dihydrogen phosphate + 0.5g magnesium sulfate + 1L water.

[0015] Preferably, the culture conditions in step S2 are: cultured in the dark at 24℃±1℃ for 8 days; and the culture conditions in step S3 are: cultured on a shaker at 120r / min at 24℃±1℃ for 5 days in the dark.

[0016] Preferably, the almond milk is prepared by grinding roasted almonds with 5-8 times their volume of water into a paste, then boiling for 20-30 minutes and filtering to remove the residue to obtain almond milk.

[0017] Preferably, the preparation method of the Buddleja officinalis salt water extract includes the following steps:

[0018] (1) Wash the whole plant of fresh Buddleja officinalis and grind it into a paste to obtain whole plant paste for later use;

[0019] (2) Add 3-5 times the volume of water to the whole plant pulp, then add sodium chloride to adjust the sodium chloride content to 10%-12%, then ultrasonically vibrate for 10-20 minutes, then press and filter to obtain the filtrate for later use.

[0020] (3) The above filtrate was spray-dried to obtain Buddleja officinalis salt extract.

[0021] Preferably, the power of the ultrasonic oscillation in step (2) is 400-600W.

[0022] Preferably, in step (3), the inlet air temperature of the spray dryer is 180°C, the outlet air temperature is 90°C, and the processing capacity is 80 kg / h.

[0023] This invention provides a method for cultivating white ginseng fungus on Simao pine stumps, which has the following advantages compared with the prior art:

[0024] Using the remaining tree stumps after logging in the Simao Pine Forest Farm as a culture medium for *Pterocarya stenoptera*, the stumps only need to be sterilized by burning in the early stage and then irrigated with a nutrient solution prepared from *Buddleja officinalis* salt water extract before the liquid inoculum of *Pterocarya stenoptera* can be directly inoculated for planting. Furthermore, using almond milk and *Buddleja officinalis* salt water extract as exogenous additives in the liquid culture medium of *Pterocarya stenoptera* can effectively promote the rapid growth of *Pterocarya stenoptera* after inoculation into the tree stumps, increase the yield of *Pterocarya stenoptera* inoculated into the tree stumps, comprehensively utilize the resources of the Simao Pine Forest Farm, and improve the economic benefits of agricultural production. Attached Figure Description

[0025] Figure 1 This is a schematic diagram illustrating the sterilization of old stumps using a blowtorch in an embodiment of the present invention;

[0026] Figure 2 This is a schematic diagram illustrating the sterilization of new piles using a blowtorch in an embodiment of the present invention;

[0027] Figure 3 This is a schematic diagram showing the growth of white ginseng fungus on the old stumps in Experiment 1 of this invention in July.

[0028] Figure 4 This is a schematic diagram showing the growth of white ginseng fungus on the old stumps of control group 1 in July in an embodiment of the present invention;

[0029] Figure 5 This is a schematic diagram showing the growth of *Panax ginseng* on the new stumps in experimental group 6 of this invention in July.

[0030] Figure 6 This is a schematic diagram showing the growth of white ginseng fungus on the new stumps in experimental group 9 of this invention in July;

[0031] Figure 7 This is a schematic diagram showing the growth of white ginseng fungus on the new stumps in experimental group 10 of this invention in July.

[0032] Figure 8 This is a schematic diagram showing the growth of *Panax ginseng* on the new stumps of control group 2 in July according to an embodiment of the present invention. Detailed Implementation

[0033] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. Example 1:

[0034] I. Preparation of Almond Milk:

[0035] Grind roasted almonds with 6 times their volume of water into a paste, then boil for 25 minutes and filter to remove the residue to obtain almond milk.

[0036] II. Preparation of Buddleja officinalis extract:

[0037] 1. Preparation of water extract of Buddleja officinalis:

[0038] (1) Wash the whole plant of fresh Buddleja officinalis and grind it into a paste to obtain whole plant paste for later use;

[0039] (2) Add 4 times the volume of water to the whole plant pulp, then ultrasonically vibrate at 400W for 15 minutes, then press and filter, and spray dry the filtrate (inlet air temperature 180℃, outlet air temperature 90℃, processing capacity 80kg / h).

[0040] 2. Preparation of Buddleja officinalis saline extract:

[0041] (1) Wash the whole plant of fresh Buddleja officinalis and grind it into a paste to obtain whole plant paste for later use;

[0042] (2) Add 4 times the volume of water to the whole plant pulp, then add sodium chloride to adjust the sodium chloride content to 11%, then ultrasonically vibrate at 400 for 15 minutes, then press and filter, and spray dry the filtrate (inlet air temperature 180℃, outlet air temperature 90℃, processing capacity 80kg / h).

[0043] 3. Preparation of Buddleja officinalis ethanol extract:

[0044] (1) Wash the whole plant of fresh Buddleja officinalis and grind it into a paste to obtain whole plant paste for later use;

[0045] (2) Add 4 times the volume of 75% ethanol solution to the whole plant pulp, then ultrasonically vibrate at 400 for 15 minutes, then press and filter, and spray dry the filtrate (inlet air temperature 180℃, outlet air temperature 90℃, processing capacity 80kg / h).

[0046] III. Preparation of Culture Media and Nutrients:

[0047] 1. Potato protein culture medium:

[0048] 200g potatoes + 20g glucose + 3g peptone + 2g potassium dihydrogen phosphate + 18g agar + 0.5g magnesium sulfate + 1L water;

[0049] 2. Potato liquid culture medium:

[0050] 200g potatoes + 20g glucose + 3g peptone + 2g potassium dihydrogen phosphate + 0.5g magnesium sulfate + 1L water;

[0051] 3. XZ-1 liquid culture medium:

[0052] Almond milk 15mL + Buddleja officinalis saline extract 700mg + potato liquid culture medium 1L;

[0053] 4. XZ-2 liquid culture medium:

[0054] Almond milk 15mL + Buddleja officinalis aqueous extract 700mg + potato liquid culture medium 1L;

[0055] 5. XZ-3 liquid culture medium:

[0056] Almond milk 15mL + Buddleja officinalis ethanol extract 700mg + potato liquid culture medium 1L;

[0057] 6. XM liquid culture medium:

[0058] 15 mL almond milk + 1 L potato broth;

[0059] 7. ZM liquid culture medium:

[0060] 700mg of Buddleja officinalis saline extract + 1L of potato liquid culture medium.

[0061] All the above culture media were sterilized by high temperature and high pressure steam (121℃, 20 min) before use.

[0062] 8. Preparation of Nutrient 1:

[0063] Prepare an aqueous solution of 300 mg / L Buddleja officinalis salt extract, and sterilize it by high temperature and high pressure steam (121℃, 20 min) to obtain nutrient solution 1.

[0064] 9. Preparation of Nutrient 2:

[0065] Prepare an aqueous solution of 300 mg / L Buddleja officinalis extract, and sterilize it by high temperature and high pressure steam (121℃, 20 min) to obtain nutrient solution 2.

[0066] 10. Preparation of Nutrient 3:

[0067] Prepare an aqueous solution of 300 mg / L Buddleja officinalis ethanol extract, and sterilize it by high temperature and high pressure steam (121℃, 20 min) to obtain nutrient solution 3.

[0068] IV. Preparation of liquid inoculum of *Pleurotus ostreatus*:

[0069] The fruiting bodies of white ginseng that grow naturally on dead trees of Pinus simaoensis in the wild were used as the strain. Rice-grain-sized pieces were taken from the junction of the fan blade and the stipe as the mother culture and inoculated into potato protein medium. The mycelium was obtained by culturing in the dark at 24℃±1℃ for 8 days.

[0070] Cut a 1cm×1cm mycelial growth block from potato protein medium, add 700μL of sterile water, and grind and homogenize using a grinder to obtain a suspension for later use.

[0071] Take 2 mL of the suspension and inoculate it onto the liquid culture medium of XZ-1, XZ-2, XZ-3, XM and ZM respectively. Incubate on a shaker at 120 r / min at 24℃±1℃ in the dark for 5 days to obtain the liquid strains of white ginseng in each group.

[0072] V. Field Planting Experiment:

[0073] The Simao pine forest farm in Jinggu Dai and Yi Autonomous County, Yunnan Province, was selected as the experimental site. Eighteen old and eighteen new Simao pine stumps were selected from the forest farm (stumps older than one year after logging were considered old stumps, and those left from logging in the current year were considered new stumps) as experimental substrates (the diameter of both old and new stumps was 15±2cm). Both old and new stumps were divided into six groups of three stumps each. Two months before the rainy season, the surface of the stumps and the surrounding area within a 20-50cm radius were sterilized by burning with a blowtorch. Figure 1 (Sterilization of old plantlets) and Figure 2 (New stump sterilization) As shown, after scorching and cooling, the stump surface was thoroughly soaked with nutrient solution. After air drying for 2 days, the corresponding liquid inoculum was poured onto the stump (200 mL per stump) on April 2, 2025. The white ginseng fungus on each stump was harvested on July 20, 2025, and the average yield of white ginseng fungus per stump was calculated, as shown in Table 1 below:

[0074] Table 1

[0075]

[0076] As shown in the table above, the use of strains cultured in XZ-1 liquid medium in combination with nutrient agent 1 can effectively increase the yield of *Pinus sylvestris* inoculated on *Pinus koraiensis* stumps, comprehensively improve the utilization rate of agricultural planting, and enhance economic benefits.

[0077] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for cultivating white ginseng fungus on Pinus simonii stumps, characterized in that, The method includes the following steps: S1. Use new or old stumps of Pinus simonii after felling as planting material. Two months before the start of the rainy season, use a blowtorch to sterilize the surface of the new or old stumps to obtain sterilized stumps for later use. S2. Select the fruiting bodies of white ginseng that grow naturally on dead trees of Pinus sylvestris in the Pinus sylvestris forest as the strain, and inoculate the strain into potato protein medium to cultivate mycelium; S3. The mycelium is mixed with sterile water and ground to prepare a suspension. The suspension is then inoculated into a liquid culture medium with the following formula: 10-20 mL almond milk + 600-800 mg Buddleja officinalis salt extract + 1 L potato liquid culture medium. This yields a liquid culture. S4. Prepare a nutrient solution of 200-400 mg / L of Buddleja officinalis salt extract. On a sunny day, thoroughly pour the sterilized nutrient solution onto the surface of the sterilized pile material once, and then let it air dry naturally to obtain the pre-treated pile material for later use. S5. Pour the liquid inoculum onto the pretreated pile material to complete the inoculation. Harvest 3-4 months later.

2. The method according to claim 1, characterized in that: In step S1, after the blowtorch sterilizes the surface of the new or old stake by burning, the vegetation around the new or old stake is also burned, with the treatment range being 20-50cm in radius of the new or old stake.

3. The method according to claim 1, characterized in that: The formula for the potato protein culture medium is 200g potato + 20g glucose + 3g peptone + 2g potassium dihydrogen phosphate + 18g agar + 0.5g magnesium sulfate + 1L water.

4. The method according to claim 1, characterized in that: The formula for the potato liquid culture medium is: 200g potato + 20g glucose + 3g peptone + 2g potassium dihydrogen phosphate + 0.5g magnesium sulfate + 1L water.

5. The method according to claim 1, characterized in that: The culture conditions in step S2 are: cultured in the dark at 24℃±1℃ for 8 days; the culture conditions in step S3 are: cultured on a shaker at 120r / min at 24℃±1℃ for 5 days in the dark.

6. The method according to claim 1, characterized in that: The almond milk preparation method is to grind roasted almonds with 5-8 times their volume of water into a paste, then boil for 20-30 minutes and filter to remove the residue to obtain almond milk.

7. The method according to claim 1, characterized in that, The preparation method of the Buddleja officinalis salt water extract includes the following steps: (1) Wash the whole plant of fresh Buddleja officinalis and grind it into a paste to obtain whole plant paste for later use; (2) Add 3-5 times the volume of water to the whole plant pulp, then add sodium chloride to adjust the sodium chloride content to 10%-12%, then ultrasonically vibrate for 10-20 minutes, then press and filter to obtain the filtrate for later use. (3) The above filtrate was spray-dried to obtain Buddleja officinalis salt extract.

8. The method according to claim 7, characterized in that: The power of the ultrasonic oscillation in step (2) is 400-600W.

9. The method according to claim 7, characterized in that: In step (3), the inlet air temperature of the spray dryer is 180℃, the outlet air temperature is 90℃, and the processing capacity is 80kg / h.

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