Fungus material for gastrodia elata cultivation and cultivation method

Abstract

The invention relates to the technical field of gastrodia elata cultivation, in particular to a fungus material for gastrodia elata cultivation and a cultivation method.The fungus material contains plants capable of inhibiting pathogenic bacteria causing gastrodia elata tuber rot disease, and the fungus material is composed of, by mass, 58% of sawdust, 30% of cottonseed hulls, 10% of maytenus stems, 1% of gypsum powder, 0.5% of KH2PO4 and 0.5% of MgSO4. The fungus material provided by the invention contains the plant maytenus stems capable of inhibiting pathogenic bacteria causing the gastrodia elata tuber rot disease, the ethanol extract of the maytenus stems can efficiently inhibit the pathogenic bacteria primary verticillium atrophaeum causing the gastrodia elata tuber rot disease, and in addition, the ethanol extract of the maytenus stems has no obvious inhibiting effect on armillaria mellea; the method ensures that a gastrodia elata-armillaria mellea symbiotic system is not interfered, the field experiment control effect is 76.85% when the fungus material is used and a method for spraying the maytenus stem ethanol extract bactericide is combined, and the method has important value for ensuring sustainable development of the gastrodia elata industry.

Description

Technical Field

[0001] The present invention relates to the technical field of Gastrodia elata cultivation, and particularly relates to a fungus log for Gastrodia elata cultivation and a cultivation method. The fungus log provided by the present invention contains plants that inhibit the pathogenic bacteria causing the rot of Gastrodia elata tubers. Background Art

[0002] Gastrodia elata Blume is a heterotrophic perennial herb. Its seeds symbiose with Mycena osmundicola, and its growth depends on Armillaria mellea to provide nutrients. Gastrodia elata is used as medicine with its dried tubers, having the effects of calming endogenous wind and stopping convulsions, suppressing hyperactive liver-yang, expelling wind and dredging collaterals, etc., and is a precious Chinese medicinal material that can be used both as medicine and food in China. Zhaotong City, Yunnan Province has become the core production area of Gastrodia elata due to its unique climate and ecological environment. However, after nearly 40 years of large-scale cultivation, the phenomenon of continuous cropping has become increasingly common, and the diseases of Gastrodia elata have become increasingly serious, resulting in reduced yields and deteriorated quality, seriously affecting the economic benefits of hemp farmers. Pathogenic microorganisms are one of the important factors leading to the incidence of diseases and reduced yields of Gastrodia elata. It is statistically shown that the incidence of the rot disease of Gastrodia elata tubers caused by pathogenic bacteria results in at least a 20% reduction in production every year, and can reach as high as 50% - 60% in severe cases, causing heavy direct economic losses and seriously threatening the sustainable development of the industry.

[0003] On the one hand, the pathogenic bacteria of Gastrodia elata infect the fungus log, compete with Armillaria mellea for nutrients, slow down the growth of Armillaria mellea, disrupt the nutritional balance between Gastrodia elata and Armillaria mellea, and reduce the yield of Gastrodia elata; on the other hand, the pathogenic bacteria directly infect the tubers of Gastrodia elata, causing the rot of Gastrodia elata tubers. There have been many research reports on symptom investigation and pathogen identification of various pathogenic bacteria causing tuber rot, such as Fusarium oxysporum F. oxysporum Schecht ), Trichoderma viride Trichodennaviride Persexfr ), Lepiota cristata Pholioiapraecoi Persexfr) , Mycogone perniciosa Mycogoneperniciosa Magn. ), Botrytis cinerea Botrytis cine⁃rea Pers.ex Fr. ), Bionectria ochroleuca I. cyclaminicola ), etc. Some research shows (Su Jianyun, Tang Jingying, Dong Xian, etc. Identification of the pathogen of Gastrodia elata tuber rot disease and determination of its pathogenicity [J]. Acta Phytopathologica Sinica, 2023, 53 (06): 1241 - 1245.) that through the isolation and identification of rotten black Gastrodia elata collected from multiple Gastrodia elata planting bases in Yunnan region, it is shown that the main pathogenic bacteria causing the rot disease of Gastrodia elata in Yunnan are Verticillium albo - atrum, which causes the vascular bundle of Gastrodia elata to turn brown and necrotic, thus leading to the rot of Gastrodia elata tubers. There are relatively many reports on the prevention and control of pathogenic bacteria such as Fusarium oxysporum and Trichoderma viride in existing research, while there are few reports on the prevention and control of Verticillium albo - atrum.

[0004] Tuber rot of Gastrodia elata is an important disease of Gastrodia elata. Its pathogenic bacteria are complex and diverse. It can not only inhibit the growth of Armillaria mellea, but also cause the rot of Gastrodia elata tubers. Traditional control methods such as using chemical fungicides like carbendazim and azoxystrobin are prone to induce drug resistance and are not conducive to the concept of high-quality green development of traditional Chinese medicine. As a new type of pesticide, botanical fungicides are gradually replacing the original chemical pesticides due to their high quality and efficiency, low toxicity and low residue, and environmental safety and friendliness, and are becoming a research hotspot for plant disease control in recent years. General fungicides only simply screen for antibacterial activity against pathogenic bacteria. However, the cultivation mode of Gastrodia elata is special. Its growth depends on Armillaria mellea to provide nutrients. Chemical fungicides may kill or inhibit Armillaria mellea, resulting in Gastrodia elata being unable to obtain sufficient nutrients from Armillaria mellea, thus affecting the growth and yield of Gastrodia elata. The purpose of this invention is to screen botanical fungicides that are safe for Armillaria mellea and can efficiently inhibit Verticillium albo-atrum, and to be used for field control of Gastrodia elata diseases, providing a more environmentally friendly and sustainable technical support for the long-term cultivation of Gastrodia elata. Summary of the Invention

[0005] The purpose of this invention is to provide a fungus material and a cultivation method for Gastrodia elata cultivation to solve the problems in the background. To achieve the above purpose, the present invention provides the following technical solutions: The present invention provides a fungus material for Gastrodia elata cultivation, and each component is calculated by mass percentage and consists of 58% wood chips, 30% cottonseed hulls, 10% Maytenus hookeri stems, 1% gypsum powder, 0.5% KH2PO4, and 0.5% MgSO4.

[0006] Furthermore, the present invention provides a preparation method of the fungus material. Take the corresponding masses of each component according to the ratio. Dissolve gypsum powder, KH2PO4, and MgSO4 in water and then add them to the mixture of wood chips, cottonseed hulls, and Maytenus hookeri stems. Add water and mix evenly to make the water content of the mixture 60% - 65%. Build a pile and ferment for 3 days, then bag, sterilize, and after cooling, inoculate with the secondary Armillaria mellea strain to prepare the medicated fungus material, and use it after culturing for 3 - 4 months.

[0007] Furthermore, the present invention provides a cultivation method for Gastrodia elata, including the following steps: (1) Prepare the medicated fungus material for standby according to the preparation method; (2) Select a plot of land, prepare the planting pits, dry them in the sun, then spray a solution of ethanol extract of Maytenus hookeri stems with a concentration of 35 g / L. After 5 days of spraying, lay the medicated fungus material and natural fungus material alternately in the planting pits with a spacing of 3 cm, and fill the spacing with new soil; (3)Prepare the seed hemp, plant the seed hemp beside the medicinal fungus logs. After arranging the seed hemp, fill the gaps with fresh soil, cover the pond, and conduct routine management. Spray the ethanol extract solution of Maytenus stem with a concentration of 35 g / L every 3 months, and apply it three times during the entire planting period. Harvest the Gastrodia elata in late November - December of the year of planting.

[0008] Furthermore, the natural fungus logs are tree species of Quercus in Fagaceae.

[0009] Furthermore, the tree species in Fagaceae include Cyclobalanopsis glauca, Quercus variabilis, or Quercus acutissima.

[0010] Furthermore, the preparation process of the ethanol extract of Maytenus stem is as follows: Take the dried Maytenus stem sample, crush it and filter it through a 40 - mesh sieve, collect the powder, weigh the dry powder at a material - to - liquid ratio of 1:10 (w / v) and add 90% ethanol solution to a container. Fix the container in an ultrasonic cleaner for ultrasonic extraction, centrifuge, and after filtration, concentrate it by rotary evaporation to obtain the ethanol extract of Maytenus stem.

[0011] Furthermore, the present invention provides the application of the ethanol extract of Maytenus stem or the ethanol extract of Ilex pubescens fruit in inhibiting Verticillium albo - atrum.

[0012] Furthermore, the present invention provides the application of the ethanol extract of Maytenus stem or the ethanol extract of Ilex pubescens fruit in preventing and controlling the rot disease of Gastrodia elata tubers.

[0013] Furthermore, the present invention provides the application of the ethanol extract of Maytenus stem or the ethanol extract of Ilex pubescens fruit in preparing a preventive and control agent for the rot disease of Gastrodia elata tubers.

[0014] Furthermore, the present invention provides a method for preventing and controlling the rot disease of Gastrodia elata tubers, applying a preparation containing the ethanol extract of Maytenus stem or the ethanol extract of Ilex pubescens fruit to the disease - occurring plots.

[0015] The technical effects achieved by the present invention are as follows: The indoor safety screening test of the present invention shows that the ethanol extracts of Maytenus stem and Ilex pubescens fruit do not damage the symbiotic system of Gastrodia elata - Armillaria mellea. The indoor control efficacy determination against the main pathogenic fungus Verticillium albo - atrum causing Gastrodia elata diseases shows that the ethanol extracts of Maytenus stem and Ilex pubescens fruit have excellent control performance against Verticillium albo - atrum. When the drug concentration is 20 mg / mL, the antibacterial rate of the ethanol extract of Ilex pubescens fruit is as high as 90.36%, and its EC 50 value is 7.67 mg / mL; the antibacterial rate of the ethanol extract of Maytenus stem is 89.32%, and the EC 50The value is 8.53 mg / mL; in the field experiment, by using the medicinal fungus materials added with the ethanol extract of Maytenus hookeri stem and the ethanol extract of Ilex pubescens fruit, and at the same time combining with the method of spraying the corresponding plant-derived fungicides, the incidence of tuber rot of Gastrodia elata was significantly reduced. While by using the method of adding 10% Maytenus hookeri stem to the fungus materials and at the same time combining with spraying the fungicide of the ethanol extract of Maytenus hookeri stem, the control effect was 76.85%. Detailed implementation manners

[0016] The present invention will be further elaborated through the following examples. It should be emphasized that these examples are only used as examples to illustrate the principles and effects of the present invention, and do not constitute a limitation to the protection scope of the present invention. In the following examples, unless otherwise specified, the experimental methods mentioned are all conventional methods commonly used in the art. Similarly, the materials, reagents, etc. used in the examples can be obtained through conventional commercial channels unless otherwise specified.

[0017] Example 1 Extraction of active ingredients from test plant materials 1. Plant materials The plant list and collection parts are shown in Table 1.

[0018] Table 1 Information of test plants Serial number Species name Latin name Family and genus Plant tissue 1 Maytenus hookeri Maytenus hookeri Loes Celastraceae, Maytenus Stem 2 Maytenus hookeri Maytenus hookeri Loes Celastraceae, Maytenus Leaf 3 Maytenus hookeri Maytenus hookeri Loes Celastraceae, Maytenus Fruit 4 Ilex pubescens Ilex pubescens Aquifoliaceae, Ilex Stem 5 Ilex pubescens Ilex pubescens Aquifoliaceae, Ilex Leaf 6 Ilex pubescens Ilex pubescens Aquifoliaceae, Ilex Fruit 7 Eleutherococcus senticosus Eleutherococcus senticosus Araliaceae, Eleutherococcus Stem 8 Eleutherococcus senticosus Eleutherococcus senticosus Araliaceae, Eleutherococcus Leaf 9 Eleutherococcus senticosus Eleutherococcus senticosus Araliaceae, Eleutherococcus Fruit 2. Extraction method The ultrasonic extraction method was adopted. First, take the dried plant samples, crush them and filter them through a 40-mesh sieve, and collect the uniform powder for standby. Then, weigh the dry powder sample at a solid-liquid ratio of 1:10 (w / v) into a conical flask, add 90% ethanol solution, fix the conical flask in an ultrasonic cleaner, set the ultrasonic power to 300 W, the frequency to 40 kHz, and extract at 40 °C for 45 min. After the extraction is completed, immediately vacuum filter with double-layer qualitative filter paper (pore size 10-12 μm) to collect the filtrate. The residue was extracted 2 times repeatedly, and the three filtrates were combined, centrifuged at 8000 r / min for 10 min at 4 °C, and the filtrate was rotary evaporated and concentrated until there was no alcohol smell to obtain the plant ethanol extract. Add 90% ethanol solution to the extract for re-dissolution to prepare a mother liquor with a concentration of 1 g / ml, and store it in a 4 °C refrigerator for standby.

[0019] Example 2 Safety screening experiment of Armillaria mellea 1. Strains and culture media Test strains: Armillaria mellea was preserved in the laboratory and identified as Armillaria gallica.

[0020] Culture medium: Potato dextrose agar (PDA) medium (200 g of potato, 20 g of glucose, 15 g of agar, made up to 1 L with distilled water, pH 6.8, sterilized at 121 °C for 20 min).

[0021] Experimental methods Activation of bacterial strains: Armillaria mellea was inoculated onto PDA plates and cultured in the dark at 25 °C for 7 days. The edge mycelial blocks were taken for standby.

[0022] Preparation of drug-containing plates: The mother liquor of the plant ethanol extract was diluted with sterile water and then mixed with the melted PDA medium (45 °C), and poured into sterile petri dishes to prepare drug-containing plates with a final concentration of 20 mg / ml.

[0023] Determination of antibacterial rate: Sterile punchers were used to take 0.6 cm diameter bacterial cakes and inoculate them in the center of the drug-containing plates, and cultured at 25 °C for 10 days. Four replicates were set for each treatment. Data were measured when the mycelia in the control group grew to 2 / 3 of the petri dish. The control group was prepared by adding an equal amount of sterile water to the melted PDA medium, preparing the plate, and inoculating the bacteria. When the control mycelia were almost full in the petri dish, the diameter (mm) of the mycelia was measured using the cross-cross method, and the antibacterial rate was calculated.

[0024]

[0025] Experimental results The antibacterial effects of the ethanol extracts of the stems, leaves and fruits of three plants on Armillaria mellea are shown in Table 2. The results show that when the extract concentration is 20 mg / ml, the ethanol extract of the fruits of Ilex pubescens shows a certain promoting effect. The antibacterial rate of the ethanol extract of the stems of Ilex pubescens is 2.02%, the antibacterial rate of the ethanol extract of the leaves of Maytenus hookeri is 0.41%, and the antibacterial rate of the ethanol extract of the stems of Maytenus hookeri is 1.15%. According to the antibacterial situation, the stems of Maytenus hookeri, the leaves of Maytenus hookeri, and the stems of Ilex pubescens with almost no inhibitory effect, as well as the fruits of Ilex pubescens with a certain promoting effect, were selected for the antibacterial experiment on Verticillium albo-atrum.

[0026] Table 2 Inhibitory effects of ethanol extracts of different parts of the tested plants on Armillaria mellea Serial number Species name Plant tissue Inhibitory rate (%) 1 Maytenus hookeri Stem 1.15 2 Maytenus hookeri Leaf 0.41 3 Maytenus hookeri Fruit 8.43 4 Ilex pubescens Stem 2.02 5 Ilex pubescens Leaf 6.47 6 Ilex pubescens Fruit -1.56 7 Eleutherococcus senticosus Stem 42.78 8 Eleutherococcus senticosus Leaf 7.12 9 Eleutherococcus senticosus Fruit 14.36 Screening of antibacterial activity of plant ethanol extracts against Verticillium albo-atrum in Example 3 1. Pathogen treatment Tested pathogen: Verticillium albo-atrum, isolated, identified and preserved in the laboratory.

[0027] Activation of the strain: Inoculate the cryopreserved strain on PDA medium (200 g of potato, 20 g of glucose, 15 g of agar, made up to 1 L with distilled water, pH 6.8), and culture it in the dark at 25°C for 7 days until the mycelium covers the plate. Pick a fresh mycelium block (5 mm in diameter) from the edge and transfer it to a fresh PDA plate, and continue to culture it at 25°C for 5 days to ensure the activity of the strain.

[0028] Preparation of spore suspension: Scrape the conidia on the surface of the medium with a sterile scalpel and place them in a sterile aqueous solution of Tween-80 with a concentration of 0.05% to prepare a spore suspension of 1×10 6 CFU / mL for standby.

[0029] Determination of the antibacterial rate and virulence of plant ethanol extracts Dilute the mother liquor of ethanol extracts from Maytenus hookeri stems, Maytenus hookeri leaves, Ilex pubescens fruits, and Ilex pubescens stems with sterile water, and use sterile PDA medium to make drug-containing plates with final concentration gradients of 5, 10, 15, and 20 mg / mL. At the same time, set a blank control containing a sterile water medium. Set 3 parallels for each concentration, and treat the blank control in the same way. Use a sterile punch to take an activated mycelium block with a diameter of 0.5 cm and inoculate it in the center of the drug-containing plate, and culture it in the dark at 25°C for 10 days to determine the antibacterial rate.

[0030] To further quantify the virulence effects of the ethanol extracts of 4 plants on Verticillium albo-atrum, we converted the concentration (mg / mL) of each drug into a logarithm value (x), and converted the average value of three replicates of the mycelial growth inhibition rate into a probit value (y). Subsequently, use statistical software to perform regression analysis on these data to fit the virulence regression equations of the ethanol extracts of the four plants on Verticillium albo-atrum. Finally, solve the equation to obtain the EC 50 value of each drug, that is, the drug concentration required to achieve a 50% growth inhibition rate. The smaller the EC 50 value, the stronger the virulence of the drug to Verticillium albo-atrum and the more significant the antibacterial effect.

[0031] Experimental results The antibacterial results and virulence of the ethanol extracts of 4 plants on Verticillium albo-atrum are shown in Table 3. As the concentration of the extract increases, the antibacterial effect becomes more obvious. When the drug concentration is 20 mg / mL, the control effects are in the order of Ilex pubescens fruits > Maytenus hookeri stems > Ilex pubescens stems > Maytenus hookeri leaves. Among them, the ethanol extract of Ilex pubescens fruits has the best inhibitory effect on Verticillium albo-atrum, with an antibacterial rate of 90.36%. Followed by Maytenus hookeri stems, with an antibacterial rate of 89.32%; Maytenus hookeri leaves have the worst inhibitory effect on Verticillium albo-atrum, with an antibacterial rate of only 62.78%. From the test results, the EC 50They are all different, and there are obvious differences in virulence. Among them, the fruits of Ilex pubescens have the strongest virulence against Verticillium albo - atrum, and the EC 50 value is 7.67 mg / mL. The stems of Maytenus hookeri come second, and the EC 50 value is 8.53 mg / mL. The toxicity of the stems of Ilex pubescens and the leaves of Maytenus hookeri decreases in turn, and the EC 50 values are 12.90 and 14.93 mg / mL respectively.

[0032] Table 3 Antibacterial effect and virulence determination of plant ethanol extracts against Verticillium albo - atrum

[0033] Example 4 Field control effect test of Gastrodia elata 1. Materials and experimental site Seed tubers: Gastrodia elata f. glauca; Armillaria mellea: Armillaria gallica; Natural fungal materials: Cyclobalanopsis glauca (diameter 4 - 8 cm, 6 - 10 small fish - scale cuts made on the wooden section); Medicated fungal materials: CK: 68% sawdust, 30% cottonseed hulls, 1% gypsum powder, 0.5% KH2PO4 and 0.5% MgSO4; T1: 68% sawdust, 30% cottonseed hulls, 1% gypsum powder, 0.5% KH2PO4 and 0.5% MgSO4; On the basis of this combination, an additional 0.05% ethanol extract of the stems of Maytenus hookeri is added; T2: 68% sawdust, 30% cottonseed hulls, 1% gypsum powder, 0.5% KH2PO4 and 0.5% MgSO4; On the basis of this combination, an additional 0.05% ethanol extract of the fruits of Ilex pubescens is added; T3: 58% sawdust, 30% cottonseed hulls, 10% stems of Maytenus hookeri (sieved through 30 - mesh sieve), 1% gypsum powder, 0.5% KH2PO4 and 0.5% MgSO4; Experimental site: Gastrodia elata planting base in Xiaocaoba, Zhaotong, Yunnan, altitude 1910 m, central coordinates (103°71′05″E, 27°35′17″N).

[0034] Preparation of medicated fungal materials Dissolve gypsum powder, KH2PO4, and MgSO4 in water, then add them to the mixture of sawdust, cottonseed hulls, or Maytenus stem. Add water and mix well to make the water content of the mixture 60% - 65%. After piling up and fermenting for 3 days, dissolve the plant ethanol extract of T2 and T3 and spray it evenly onto the mixture. For CK, add an equal amount of water. Put the mixed product into polyethylene plastic bags, 1 kg per bag; tie the bag mouths tightly; under 121 °C, sterilize under high pressure for 1 - 2 h; after sterilization and cooling to room temperature, transport it to the sterile room, inoculate with the secondary Armillaria mellea strain, and transport it to the culture room for cultivation, cultivate at 25 °C for 3 - 4 months.

[0035] Field experiment method Select a replanted Gastrodia elata plot where tuber rot occurred in the first crop. Level the replanted planting pits (60 cm × 50 cm × 30 cm), and after sunning for 20 days, spray 1 L of a plant ethanol extract solution with a concentration of 35 g / L onto each planting pit. For the control, use an equal amount of water for treatment. After 5 days of spraying, lay the medicated fungus materials and natural fungus materials crosswise in the planting pits, with a spacing of about 3 cm, and fill the spacing with new soil; select about 30 g of white-headed Gastrodia elata as the seed tubers; plant the seed tubers beside the medicated fungus materials, with a distance of 10 cm between the seed tubers. Plant 5 seed tubers beside each row of medicated fungus materials, with a total of 20 seed tubers placed. Set 5 replicates for each treatment. After placing the seed tubers, fill the gaps with soil, cover the pits, and conduct routine management. Spray 1 L of a plant ethanol extract solution with a concentration of 35 g / L every 3 months, and apply it three times during the entire planting period; plant at the beginning of the test year and harvest in November at the end of the year. When the farmers harvest Gastrodia elata, investigate the incidence of Gastrodia elata diseases, put the collected Gastrodia elata samples into self-sealing bags and bring them back to the laboratory, and statistically analyze the healthy and diseased Gastrodia elata for each treatment; and calculate the disease index and control effect according to the Gastrodia elata tuber disease grading standard.

[0036] Table 4 Gastrodia elata tuber disease grading standard

[0037]

[0038] Table 5 Field disease control effect Treatment Incidence rate (%) Disease index Control effect (%) CK 50 27.78 - T1 14.29 10.40 62.55 T2 14.29 13.98 49.69 T3 6.67 6.43 76.85 A control effect test was carried out in the Zhaotong Gastrodia elata planting base. As shown in Table 5, it can be seen that applying the ethanol extract of Maytenus stem and the ethanol extract of Ilex pubescens fruits can significantly reduce the incidence of Gastrodia elata tuber rot when planting Gastrodia elata in replanted plots, indicating that the plant-derived fungicide provided by the present invention has good control effects. Among them, the control effects of adding the ethanol extract of Maytenus stem and the ethanol extract of Ilex pubescens fruits into the fungus materials are 62.55% and 49.69% respectively, and the control effect of adding Maytenus stem into the fungus materials is 76.85%, and the control effect is significant.

[0039] In summary, through systematic screening and evaluation, the present invention has identified three plants: Maytenus hookeri, Ilex pubescens, and Eleutherococcus senticosus, and determined the safety of ethanol extracts from different parts of these plants against Armillaria mellea and their inhibitory effects on Verticillium albo-atrum. Based on the safety screening results, ethanol extracts from the stems and leaves of Maytenus hookeri, the fruits and stems of Ilex pubescens were selected for the antibacterial experiment against Verticillium albo-atrum. In the antibacterial experiment against Verticillium albo-atrum, when the concentration reached 20 mg / mL, the ethanol extract from the fruits of Ilex pubescens showed the best antibacterial effect, with an antibacterial rate as high as 90.36%, and its EC 50 value was 7.67 mg / mL, significantly better than other extracts. The ethanol extract from the stems of Maytenus hookeri ranked second, with an antibacterial rate of 89.32% and an EC 50 value of 8.53 mg / mL. In the field experiment, the use of medicinal fungus materials containing ethanol extracts from the stems of Maytenus hookeri and the fruits of Ilex pubescens, combined with the spraying of corresponding plant-derived fungicides, significantly reduced the incidence of rotten Gastrodia elata tubers, with control effects of 62.55% and 49.69% respectively. When 10% of the stems of Maytenus hookeri were added to the fungus materials and combined with the spraying of the ethanol extract fungicide from the stems of Maytenus hookeri, the control effect was 76.85%. There were extremely significant differences in the control effects of different treatments. It is speculated that the stability of plant ethanol extracts is affected by high temperature or the environment, and the dosage form can be further optimized in the later stage to improve the effect.

[0040] In conclusion, the ethanol extracts from the fruits of Ilex pubescens and the stems of Maytenus hookeri have both high safety against Armillaria mellea and high inhibitory effects on Verticillium albo-atrum, and can be used as preferred plant-derived fungicides for the green prevention and control of rotten Gastrodia elata tubers. This study provides a theoretical basis and technical support for the sustainable prevention and control of Gastrodia elata diseases, and at the same time lays an important foundation for the development and application of plant-derived fungicides.

[0041] The above are only the preferred embodiments of the present invention, and do not limit the patent scope of the present invention accordingly. Any equivalent transformation made using the content of the specification of the present invention under the concept of the present invention, or any direct / indirect application in other related technical fields, is included in the patent protection scope of the present invention.

Claims

1. A fungus material for Gastrodia elata cultivation, characterized in that, Each component is composed of 58% sawdust, 30% cottonseed hulls, 10% Maytenus stem, 1% gypsum powder, 0.5% KH2PO4, and 0.5% MgSO4 by mass percentage.

2. A preparation method of the fungus material according to claim 1, characterized in that, Take corresponding masses of each component according to the ratio. Dissolve gypsum powder, KH2PO4, and MgSO4 in water and then add them to the mixture of sawdust, cottonseed hulls, and Maytenus stem. Add water and mix evenly to make the water content of the mixture 60% - 65%. Build a pile and ferment for 3 days, then bag, sterilize, and after cooling, inoculate with the secondary Armillaria mellea strain to prepare the medicinal fungus material, which can be used after culturing for 3 - 4 months.

3. A Gastrodia elata cultivation method, characterized in that, It includes the following steps: (1) Prepare the medicinal fungus material for standby according to the preparation method of claim 2; (2) Select a site, arrange the planting pits, and after sun-drying, spray the ethanol extract solution of Maytenus stem with a concentration of 35 g / L. After 5 days of spraying, lay the medicinal fungus material and natural fungus material crosswise in the planting pits with a spacing of 3 cm, and fill the spacing with new soil; (3) Prepare the seed tubers, plant the seed tubers beside the medicinal fungus material. After placing the seed tubers well, fill the gaps with new soil, cover the pits, and conduct conventional management. Spray the ethanol extract solution of Maytenus stem with a concentration of 35 g / L every 3 months, and apply it three times during the entire planting period. Dig the Gastrodia elata at the end of November - December of the planting year.

4. The cultivation method according to claim 3, characterized in that, The natural fungus material is a tree species of the genus Quercus in the family Fagaceae.

5. The cultivation method according to claim 4, characterized in that, The tree species of the family Fagaceae include Cyclobalanopsis glauca, Quercus variabilis, or Quercus acutissima.

6. The cultivation method according to any one of claims 3 - 5, characterized in that, The preparation process of the ethanol extract of Maytenus stem is as follows: Take the dried Maytenus stem sample, crush it and filter it through a 40-mesh sieve, collect the powder, weigh the dry powder at a material-liquid ratio of 1:10 (w / v) and add 90% ethanol solution to a container. Fix the container in an ultrasonic cleaner for ultrasonic extraction, centrifuge, and after filtration, concentrate it by rotary evaporation to obtain the ethanol extract of Maytenus stem.

7. Application of ethanol extract of Maytenus hookeri stem or ethanol extract of Ilex pubescens fruit in inhibiting Verticillium albo - atrum.

8. Application of ethanol extract of Maytenus hookeri stem or ethanol extract of Ilex pubescens fruit in preventing and controlling the rot disease of Gastrodia elata tuber.

9. Application of ethanol extract of Maytenus hookeri stem or ethanol extract of Ilex pubescens fruit in preparing a preventive and control agent for the rot disease of Gastrodia elata tuber.

10. A method for preventing and controlling the rot disease of Gastrodia elata tuber, characterized in that, Apply the preparation containing the ethanol extract of Maytenus stem or the ethanol extract of Ilex pubescens fruits to the disease-occurring plots.

Images