Anoectochilus formosanus culture medium and culture method thereof

Abstract

The invention provides an anoectochilus formosanus culture medium and a culture method thereof.The anoectochilus formosanus culture medium comprises gravels, granite, perlite, bark, river sand and peat soil, and the anoectochilus formosanus culture medium is matched with the anoectochilus formosanus culture method. After anoectochilus formosanus seedlings are transplanted into the culture medium, the anoectochilus formosanus can rapidly grow in a short time by adjusting the humidity of the culture medium, the humidity and temperature of the indoor environment, the illumination duration, the brightness and the irradiation mode, the survival rate is high, and the content of effective components of adult plants is high. The anoectochilus formosanus culture medium provided by the invention is simple in formula and low in cost, and large-scale factory cultivation of anoectochilus formosanus can be met by matching with the anoectochilus formosanus culture method provided by the invention.

Description

Technical Field

[0001] The invention belongs to the technical field of agriculture, and in particular relates to a roxburghii roxburghii cultivation medium and a cultivation method thereof. Background Art

[0002] Anoectochilus roxburghii, also known as golden thread orchid, likes moisture. Wild Anoectochilus roxburghii is suitable for growing in cool forests, rich soil humus, and excellent water and humidity conditions. It is commonly found under evergreen broad-leaved forests or in humid valleys at an altitude of 50-1600 meters, and on the dead branches and leaves layer under subtropical evergreen broad-leaved forests, mixed coniferous and broad-leaved forests or bamboo forests with complete community structures, or on humus soil between shady and wet stones. The whole plant of Anoectochilus roxburghii can be used as a Chinese medicinal material, mainly for treating tuberculosis hemoptysis, diabetes, nephritis, myasthenia gravis and other diseases; its plant shape is small and beautiful, and its leaves are beautiful. It is one of the ornamental leaf treasures with ornamental benefits indoors. It is widely used in the fields of medicine, health care, beauty and drinking products, and has broad market prospects.

[0003] Because the seeds of golden thread vine are small, the germination rate is low under natural conditions, the plants are short and grow slowly, and the degradation of forest ecosystem functions and excessive exploitation by humans have led to a continuous reduction in its wild resources. Golden thread vine has strict requirements for the growth environment, including soil, water, light, temperature and other conditions, so artificial cultivation of golden thread vine requires extremely high technical requirements. If the cultivation method is not up to standard, it will cause problems such as slow growth of golden thread vine, more diseases and pests, low survival rate and insufficient weight of mature plants.

[0004] The existing method for cultivating golden thread vine only optimizes the seedling breeding process of golden thread vine or optimizes the device for cultivating golden thread vine, but does not optimize the growth of golden thread vine from seedling to adult stage. It only selects the best solution by simply combining the cultivation medium and growth conditions of golden thread vine, but does not take into account that golden thread vine likes to grow in weakly acidic soil and its root system has extremely high requirements for air permeability and water absorption. Since golden thread vine in the natural environment likes shade and the plant is short, it is shielded by other plants and receives less sunlight, and the content of effective ingredients synthesized by photosynthesis is low. The existing method for cultivating golden thread vine does not optimize this point to increase its effective ingredient content.

[0005] Therefore, it is urgent to find a kind of roxburghii cultivation medium and its culture method to solve the above problems. Summary of the invention

[0006] The present invention provides a kind of golden thread vine cultivation matrix and its cultivation method, the matrix includes gravel, granite, perlite, bark, river sand, peat soil, the cultivation matrix cooperates with the golden thread vine cultivation method provided by the present invention, after the golden thread vine seedling is transplanted into the cultivation matrix, by adjusting the humidity of the cultivation matrix and the humidity, temperature of the indoor environment and the duration, brightness and irradiation mode of the light, the golden thread vine can be allowed to grow rapidly in a short time, the survival rate is high, and the content of active ingredients in mature plants is high. The golden thread vine cultivation matrix provided by the present invention is simple in formula and low in cost, and the golden thread vine cultivation method provided by the present invention can meet the large-scale factory cultivation of golden thread vine.

[0007] On the one hand, a golden thread vine cultivation medium includes gravel, granite, perlite, bark, river sand, and peat soil.

[0008] Gravel and granite improve the soil matrix structure. The pore structure of gravel increases the aeration and water retention of the soil matrix, which is conducive to the development of the root system of golden thread vine and can provide a good growth environment for golden thread vine. Gravel and granite are rich in minerals, such as calcium, magnesium, iron, etc. The supplementation of these minerals can accelerate the growth of golden thread vine. Adding peat soil can also reduce the occurrence of pests and diseases, because its physical properties can prevent the breeding of some pests and diseases.

[0009] The porous structure of perlite can make the soil matrix not only have drainage permeability and excellent water absorption, preventing soil compaction, but also allow the roots of golden thread vine to grasp it and promote the growth of plant roots. In addition, perlite is weakly acidic, and it can also play a role in regulating the pH of the soil, which is very beneficial for plants such as golden thread vine that like to grow in acidic soil.

[0010] The bark improves the soil matrix structure, enhances the water permeability and air permeability of the soil matrix, increases the looseness of the soil matrix, reduces the compaction of the soil matrix, and is beneficial to the growth of the golden thread vine root system; the bark contains rich organic matter and trace elements, and after fermentation treatment, it can improve the fertility of the soil matrix. For example, the humic acid in the bark can promote the growth of the golden thread vine root system and the ability to absorb nutrients, thereby improving the plant's disease resistance and yield.

[0011] River sand increases soil permeability and water retention, improves soil matrix texture, and enhances soil matrix drought resistance, which is beneficial to the growth of golden thread vine roots; river sand can regulate soil matrix temperature and prevent the soil matrix from overheating or overcooling; the particle structure of river sand helps prevent soil compaction and keeps the soil loose; in addition, river sand is rich in minerals and trace elements, which can increase the fertility of the soil matrix and promote the growth and development of golden thread vine.

[0012] Peat soil has good water retention properties, can absorb and retain a large amount of water, provide a continuous water source for golden thread vine, and reduce water evaporation; the loose texture and porous structure of peat soil make it have good air permeability, which is beneficial to the respiration and gas exchange of the golden thread vine root system, and promotes its growth and development; peat soil is rich in organic matter, such as nitrogen, phosphorus, potassium and other nutrients, which provide a continuous and stable supply of nutrients for golden thread vine; golden thread vine is suitable for rooting in slightly acidic soil conditions, and peat soil is usually slightly acidic, which is beneficial to the growth and development of the golden thread vine root system.

[0013] Furthermore, the mixing mass ratio of the gravel, granite, perlite, bark, river sand and peat soil is (2.4-3.6): (2.4-3.6): (0.8-1.2): (1.6-2.4): (0.8-1.2): (8-12).

[0014] Furthermore, the diameter of the gravel and granite is 1-5 cm, the diameter of the perlite is 0.3-3 mm, the length of the bark is 1-3 cm, the diameter of the river sand is 0-0.5 mm, and the thickness of the peat soil is 20-40 mm.

[0015] There was no significant difference in the cultivation medium made of gravel, granite, perlite within the above diameter range and peat soil within the above thickness range.

[0016] In some embodiments, the length of the bark is preferably 2 cm.

[0017] Furthermore, a method for cultivating golden thread lotus adopts the above-mentioned golden thread lotus cultivation medium, and the cultivation method comprises the following steps:

[0018] (1) Sterilizing the cultivation medium;

[0019] (2) Transplanting the roxburghii seedlings into the cultivation medium and cultivating them in an indoor environment;

[0020] (3) Adjust the humidity of the cultivation medium and the humidity and temperature of the indoor environment, as well as the duration, brightness, and illumination method of the lighting.

[0021] Furthermore, the sterilization method is 121° C., high pressure sterilization for 15-20 minutes.

[0022] Through high temperature and high pressure sterilization treatment, pathogenic microorganisms in the cultivation matrix, such as bacteria, fungi and viruses, can be effectively killed. On the one hand, some pathogenic microorganisms can be transmitted to crops through the cultivation matrix, causing soil-borne diseases. Sterilization treatment can inhibit the reproduction and spread of these soil-borne pathogens, thereby reducing the occurrence of golden thread vine diseases and reducing its losses caused by diseases; on the other hand, harmful microorganisms may hinder the normal growth of golden thread vine. Sterilization treatment can provide a healthy growth environment for golden thread vine and promote its growth and development; on the other hand, harmful microorganisms may destroy the structure of the cultivation matrix and reduce the quality of the cultivation matrix. Sterilization treatment can maintain the quality of the cultivation matrix and improve the fertility and water retention capacity of the cultivation matrix.

[0023] Furthermore, the humidity of the cultivation substrate is maintained at 40%-50%, the air humidity is maintained at 65%-70%, and the air humidity is maintained at 65%-70%.

[0024] Golden thread vine has extremely high requirements for water. The cultivation medium should be kept moist, but not waterlogged. If the moisture content of the cultivation medium is lower than 40%, the golden thread vine will lack water and fall over. If it is higher than 50%, the roots are prone to rot. The water used should be purified water or tap water after exposure to the sun. The water temperature should be controlled below 25°C, 23°C is the best. If the water temperature exceeds 25°C or is lower than 18°C, it will cause high pressure on the water treatment system and increase energy consumption. Golden thread vine likes a humid environment and requires the air humidity to be kept at a high level.

[0025] Furthermore, within one day, the illumination duration is 18 hours, the light brightness is maintained within 2000Lux, the illumination mode is oblique or diffuse reflection, and the temperature is controlled at 29°C-30°C; the no-light duration is 6 hours, and the temperature is controlled at 18°C-23°C.

[0026] If you set a low temperature in a bright environment and a high temperature in a dark environment, the golden thread vine will become dehydrated and fall over.

[0027] The active ingredients of Anoectochilus roxburghii include Anoectochilus roxburghii polysaccharide and Anoectochilus glycoside. The content of Anoectochilus roxburghii polysaccharide and Anoectochilus glycoside is inversely proportional to the length of dark treatment time. Too long dark treatment time will lead to a decrease in the content of active ingredients in Anoectochilus roxburghii. Therefore, light treatment simulates the longest illumination time of 18 hours for Anoectochilus roxburghii in the natural environment, which can increase the biomass of Anoectochilus roxburghii with maximum efficiency. The photosynthesis saturation point of Anoectochilus roxburghii is low, and the light intensity needs to be controlled within 2000Lux, but not less than 20Lux. If the light intensity is lower than 20Lux, Anoectochilus roxburghii may stop synthesizing active ingredients. Anoectochilus roxburghii grown in the natural environment does not like light and prefers a cool environment. Therefore, the indoor light source is refracted or diffusely reflected to the plant through a reflector to avoid direct sunlight. Too strong light intensity or direct light source will burn Anoectochilus roxburghii.

[0028] In some embodiments, the light intensity is preferably 500 Lux.

[0029] In another aspect, a substrate composition is provided for increasing the biomass of roxburghii roxburghii, wherein the substrate composition comprises gravel, granite, perlite, bark, river sand, and peat soil.

[0030] In another aspect, a substrate composition is used for improving the growth of golden thread lotus and accumulating the content of roxburghii polysaccharide and roxburghii glycoside. The substrate composition is gravel, granite, perlite, bark, river sand, and peat soil.

[0031] The present invention has the following beneficial effects:

[0032] (1) The present invention provides a culture medium for golden thread vine, the medium comprising gravel, granite, perlite, bark, river sand, and peat soil;

[0033] (2) The present invention also provides a method for cultivating Anoectochilus roxburghii. After transplanting Anoectochilus roxburghii seedlings into the cultivation medium, the humidity of the cultivation medium and the humidity, temperature, and duration, brightness, and mode of the indoor environment are adjusted, so that Anoectochilus roxburghii can grow rapidly in a short period of time, with a high survival rate and a high content of effective ingredients in mature plants.

[0034] (3) The roxburghii cultivation substrate provided by the present invention has a simple formula and low cost (reducing the amount of peat soil by 30%-50%, and the stone can be used permanently). In combination with the roxburghii cultivation method provided by the present invention, it can meet the needs of large-scale factory cultivation of roxburghii. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 The cultivation conditions of the 3-month-old golden thread vine seedlings after being transplanted into the planting tray in Example 1;

[0036] Figure 2 Combination 2 of Example 2 is a peat soil cultivation medium mainly composed of pebbles (round gravel);

[0037] Figure 3 Combination 5 of Example 2 is a peat soil cultivation medium mainly composed of gravel;

[0038] Figure 4 The peat soil cultivation matrix of combination 6 of Example 2 is mainly composed of granite. DETAILED DESCRIPTION

[0039] The present invention will be further described in detail below in conjunction with the embodiments of the accompanying drawings. It should be pointed out that the embodiments described below are intended to facilitate the understanding of the present invention and do not have any limiting effect on the present invention.

[0040] Example 1: A method for preparing a culture medium for golden thread lily and a method for cultivating it

[0041] 1. Prepare the cultivation medium: Sterilize gravel (3.5cm in diameter), granite (3.5cm in diameter), perlite (2mm in diameter), bark (2cm in length), river sand (0.2mm in diameter), and peat soil (30mm in thickness) separately at 121°C and high pressure sterilization for 15-20 minutes. Sterile peat soil can also be used directly. Then, mix the sterile gravel, granite, perlite, bark, river sand, and peat soil in a mass ratio of 3:3:1:2:1:10 and evenly pack them into the planting tray.

[0042] 2. Take out the 3-month-old golden thread lotus tissue culture seedlings from the tissue culture medium, treat them with ultrasound for 1 minute, wash them with clean water 3 times, and then transplant them into the planting tray. Cultivate the planting tray in an indoor environment, such as Figure 1 As shown;

[0043] 3. Control culture conditions

[0044] (1) Use 23°C pure water or tap water after exposure to the sun to water the cultivation medium every day to keep the moisture content of the cultivation medium at 40%-50%;

[0045] (2) Indoor humidity is maintained at 65%-70%;

[0046] (3) Within one day, the illumination time is 18 hours, using LED full light source, the light intensity is kept within 20-500 Lux, the light source is refracted onto the golden thread vine plant through a reflector to avoid direct sunlight, and the room temperature is controlled at 29℃-30℃; the time without light is 6 hours, and the room temperature is controlled at 18℃-23℃;

[0047] (4) The culture period is 90 days.

[0048] Example 2: Effects of different cultivation substrate combinations on the growth of Anoectochilus roxburghii

[0049] The difference between this embodiment and embodiment 1 is that cultivation substrates of different compositions are prepared, as shown in the following Table 1.

[0050] Table 1. Cultivation substrates of different compositions

[0051] combination Cultivation substrates of different compositions and their mass ratios one Gravel: granite: perlite: bark: river sand: peat soil = 3:3:1:2:1:10 two Pebbles (round gravel): perlite: bark: river sand: peat soil = 3:1:2:1:10 three Planting stone: perlite: bark: river sand: peat soil = 3:1:2:1:10 Four Vermiculite: perlite: bark: river sand: peat soil = 3:1:2:1:10 five Gravel: Perlite: Bark: River Sand: Peat Soil = 3:1:2:1:10 six Granite: Perlite: Bark: River Sand: Peat Soil = 3:1:2:1:10 seven Gravel: granite: bark: river sand: peat soil = 3:3:2:1:10 eight Gravel: granite: perlite: river sand: peat soil = 3:3:1:1:10 Nine Gravel: granite: perlite: bark: peat soil = 3:3:1:2:10

[0052] Combination of two cultivation media such as Figure 2 As shown, the combination of five cultivation substrates is as follows Figure 3 As shown, the combination of six cultivation substrates is as follows Figure 4 shown.

[0053] The nine groups of cultivation substrates with different compositions were planted and cultivated with Anoectochilus roxburghii according to the cultivation substrate preparation method and the Anoectochilus roxburghii cultivation method provided in Example 1. Thirty well-developed Anoectochilus roxburghii seedlings were planted in each cultivation substrate. The average fresh weight growth rate (compared with the previous fresh weight) and the survival rate of each group of Anoectochilus roxburghii were calculated every 15 days. The fresh weight growth rate of Anoectochilus roxburghii is shown in Table 2 below, and the mortality rate is shown in Table 3 below. After 90 days, the proportions of the active ingredients Anoectochilus roxburghii polysaccharide and Anoectochilus roxburghii in the fresh weight of Anoectochilus roxburghii planted in different groups of cultivation substrates were detected.

[0054] The detection methods of Anoectochilus roxburghii polysaccharide and Anoectochilus roxburghii glycoside are as follows:

[0055] All the Anoectochilus roxburghii plants in each group were equally divided into two parts, one for detecting the content of Anoectochilus roxburghii polysaccharides and the other for detecting the content of Anoectochilus roxburghii glycosides.

[0056] 1. Determination of polysaccharide content in Anoectochilus roxburghii

[0057] Take a portion of Anoectochilus roxburghii from each group and weigh it as W1.

[0058] Detection of Anoectochilus roxburghii polysaccharide by phenol-sulfuric acid method:

[0059] (1) Extraction of crude polysaccharides from Anoectochilus roxburghii: The whole herb of Anoectochilus roxburghii was dried in the sun, vacuum dried at 40°C for 1 h, crushed, and passed through a 380 μm sieve; water was added at a material-liquid ratio of 1:20, and microwave-ultrasound synergistic extraction was performed at 50°C for 30 min at 4200 r·min -1 Centrifuge for 20 minutes, take the supernatant, add water to the residue at a solid-liquid ratio of 3:20, repeat twice, combine the supernatants, concentrate the supernatant volume to 1 / 10 of the original, add Sevag reagent (chloroform: n-butanol = 4:1) to the supernatant, oscillate and centrifuge for 15 minutes, remove the middle protein layer, detect at an ultraviolet wavelength of 250-280nm until there is no absorption peak, add 3 times the volume of anhydrous ethanol, remove the supernatant, collect the precipitate, and freeze-dry for 48 hours to obtain the crude polysaccharide of Anoectochilus roxburghii, weighed as W2.

[0060] (2) Preparation of 5% phenol solution: 100.0 g of phenol was weighed by electronic balance and dissolved in a beaker, 0.10 g of aluminum sheet and 0.05 g of sodium bicarbonate were added, and the mixture was heated for distillation. 10.00 g of the fraction was collected, 190 mL of water was added to dissolve the fraction, and the fraction was placed in a brown bottle for later use;

[0061] (3) Preparation of glucose standard solution: Accurately weigh 0.1 g of glucose standard dried to constant weight at 105°C, dissolve in distilled water, and prepare 0.10 mg / mL glucose standard solution. Store at 4°C for later use.

[0062] (4) Determine the measurement wavelength: Accurately pipette 1.00 mL of 0.10 mg / mL glucose reference substance into a test tube, add 0.60 mL of 5% phenol, quickly add 6.0 mL of concentrated sulfuric acid, scan in the wavelength range of 200 to 800 nm, and determine that the maximum absorption wavelength is 490 nm.

[0063] (5) Draw a glucose standard curve: Accurately pipette 0.00, 0.20, 0.40, 0.60, 0.80, and 1.00 mL of 0.10 mg / mL glucose standard solution into a 20 mL stoppered test tube, fill with distilled water to 1.00 mL, add 0.6 mL of 5% phenol and 6.0 mL of concentrated sulfuric acid, shake well and cool, place at room temperature for 20 min, use the tube without glucose standard solution as a blank reference, and measure the absorbance at 490 nm. Draw the standard curve equation with glucose concentration X (mg / mL) as the horizontal axis and absorbance value Y as the vertical axis.

[0064] (6) Weigh 0.1 mg of the crude polysaccharide of Anoectochilus roxburghii prepared in the above step (1) into a 20 mL stoppered test tube, add 1 mL of distilled water to dissolve, then add 0.6 mL of 5% phenol and 6.0 mL of concentrated sulfuric acid, shake well and cool, and place at room temperature for 20 min. Use the tube without the test sample as a blank reference, measure the absorbance value at 490 nm, calculate the Anoectochilus roxburghii concentration of different groups of Anoectochilus roxburghii according to the standard curve, and calculate the total content W3 of Anoectochilus roxburghii polysaccharide in the crude polysaccharide W2 according to the concentration. The proportion of Anoectochilus roxburghii polysaccharide content in the fresh weight of Anoectochilus roxburghii = W3 / W1.

[0065] 2. Determination of Anoectochilus glycoside content

[0066] Take another portion of Anoectochilus roxburghii from each group and weigh it as W4.

[0067] (1) Preparation of sample solution: Dry and crush another portion of A. roxburghii in each group at 60°C, pass through a 250 μm sieve to obtain A. roxburghii powder, weigh the total weight as W5, take 0.5 g of A. roxburghii powder into a conical flask, accurately add 100 mL of methanol, seal the flask, weigh the weight, perform ultrasonic treatment (power 250 W, frequency 50 kHz) for 40 min, cool, weigh the weight again, make up the weight loss with methanol, shake well, and filter using a 0.22 μm filter membrane.

[0068] (2) HPLC-ELSD detection: Chromatographic conditions: phenomenex NH2 column (250 mm × 4.6 mm, 5 μm); acetonitrile-water (92:8, V / V) mobile phase, isocratic elution; flow rate 0.8 mL / min; column temperature room temperature; evaporative light scattering detector nebulizer chamber temperature 70 °C, carrier gas high-purity nitrogen, carrier gas flow rate 1.5 L / min; injection volume 10 μL, measured using ELSD detector.

[0069] (3) Preparation of reference solution and standard curve:

[0070] Accurately weigh 12.67 mg of roxburghii glycoside reference substance into a 5 mL volumetric flask, add methanol to the scale, and dissolve by ultrasonication to prepare a roxburghii glycoside reference substance solution containing 2.432 mg per 1 mL.

[0071] The reference solution was diluted to 2432.64, 1216.32, 304.08, 152.04, and 76.02 μg / mL, 10 μL was injected, the peak area was determined, and linear regression was performed with the logarithm of the peak area integral value (A) as the ordinate (y) and the logarithm of the reference concentration B as the coordinate (X) to obtain the regression equation, correlation coefficient and linear range of anoectochiloside.

[0072] (4) Take 10 μL of each sample solution prepared in step (1) and perform parameter detection according to the parameters provided in step (2), calculate the concentration according to the standard curve, and then calculate the total content W6 of anoectochiloside in W5 according to the concentration. The proportion of anoectochiloside in the fresh weight of Anoectochiloside = W6 / W4.

[0073] The content proportions of two active ingredients, roxburghii polysaccharide and roxburghii glycoside, in the fresh weight of roxburghii in different groups are shown in Table 4.

[0074] Table 2. Growth rate of fresh weight of Anoectochilus roxburghii planted in different crushed stone-prepared cultivation substrates

[0075] Group 15d 30d 45d 60d 75d 90d one 14％ 27％ 40％ 49％ 65％ 73％ two 14％ 22％ 35％ 42％ 53％ 65％ three 11％ 14％ 17％ 24％ 38％ 42％ Four 12％ 18％ 25％ 34％ 46％ 52％ five 14％ 20％ 28％ 36％ 42％ 55％ six 11％ 17％ 22％ 29％ 34％ 46％ seven 13％ 16％ 20％ 27％ 36％ 41％ eight 9％ 12％ 16％ 19％ 24％ 29％ Nine 10％ 15％ 18％ 23％ 27％ 32％

[0076] Table 3. Survival rate of Anoectochilus roxburghii planted in different crushed stone culture mediums

[0077]

[0078]

[0079] Table 4. Content of effective ingredients in the cultivation medium prepared with different crushed stones for planting Anoectochilus roxburghii

[0080] Active substance content Group 1 Group 2 Group 3 Group 4 Group 5 Group 6 Group 7 Group 8 Group 9 Anoectochilus polysaccharide 34.7％ 25.2％ 20.4％ 19.5％ 24.7％ 23.9％ 21.7％ 17.3％ 18.5％ Anoectochilus glycoside 24.3％ 19.6％ 17.0％ 16.7％ 15.8％ 18.4％ 16.3％ 15.7％ 15.1％

[0081] According to the data analysis in Table 2 and Table 3, the fresh weight growth rate and survival rate of the golden thread vine of combination 1 to combination 4 were compared. The difference between combination 1 and combination 2 to 4 is that combination 2 to 4 replaced the gravel and granite combination of combination 1 with single pebble (round gravel), planting stone and vermiculite respectively. It was found that the golden thread vine cultivated in combination 1 had the largest fresh weight growth and the highest survival rate within 90 days. The survival rate of the golden thread vine of combination 1 could reach 96.7% on the 90th day, while the survival rates of the other three groups were all less than 90% on the 90th day.

[0082] Comparing the fresh weight growth rate and survival rate of the golden thread vine in combination one and combinations five to nine, the difference between combination one and combinations five to nine is that the five combinations five to nine lack one of the gravel, granite, perlite, bark and river sand in combination one, and it was found that the golden thread vine cultivated in combination one had the largest fresh weight growth and the highest survival rate within 90 days, and the survival rate of the golden thread vine in combinations five to nine was less than 80% on the ninetieth day, which was much lower than the survival rate of the golden thread vine in combination one, indicating that the lack of any one of the gravel, granite, perlite, bark and river sand in combination one will reduce the fresh weight growth and survival rate of the cultivated golden thread vine.

[0083] According to the data analysis in Table 4, the content of roxburghii polysaccharide and roxburghii glycoside in the roxburghii cultivated in the cultivation medium of group 1 were significantly higher than those in the other groups. The content of roxburghii polysaccharide in the roxburghii cultivated under natural growth conditions was between 17% and 26%, and the content of roxburghii glycoside was between 15% and 17%. The roxburghii cultivated in group 1 could make the content of effective ingredients in roxburghii far exceed the content of effective ingredients in the roxburghii cultivated under natural growth conditions.

[0084] Gravel, granite, perlite, bark, and river sand can improve the structure of the cultivation substrate, increase the air permeability and water retention of the cultivation substrate, prevent soil compaction, help the development of the root system of golden thread vine, and provide a good growth environment for golden thread vine. Gravel, granite, perlite, and river sand are not only rich in minerals and trace elements, such as calcium, magnesium, and iron, which are conducive to the growth of golden thread vine, but their physical properties can also prevent the breeding of some diseases and insect pests. The porous structure of perlite allows the root system of golden thread vine to grasp it and promote the growth of plant roots. The bark is rich in organic matter and trace elements. After fermentation, it can improve the fertility of the cultivation substrate. For example, the humic acid in the bark can enhance the ability of the root system of golden thread vine to absorb, and improve the disease resistance and yield of the plant. River sand can adjust the temperature of the cultivation substrate to prevent the cultivation substrate from overheating or overcooling. Peat soil has good water retention properties, can absorb and retain a large amount of water, provide a continuous water source for golden thread vine, and reduce water evaporation; the loose texture and porous structure of peat soil make it have good ventilation, which is conducive to the respiration and gas exchange of the root system of golden thread vine, and promotes its growth and development; peat soil is also rich in organic matter, such as nitrogen, phosphorus, potassium and other nutrients, which provide a continuous and stable supply of nutrition for golden thread vine. Golden thread vine is suitable for rooting in slightly acidic soil conditions. Peat soil and perlite are usually slightly acidic, and can also play a role in regulating the pH value of the cultivation matrix. It is very beneficial for plants such as golden thread vine that like to grow in acidic soil, and is conducive to the growth and development of the root system of golden thread vine.

[0085] Based on the above analysis, gravel, granite, perlite, bark, river sand and peat soil are preferred as the cultivation medium for growing golden thread vine.

[0086] Example 3: Effect of bark length on the growth of Anoectochilus roxburghii

[0087] The difference between this embodiment and embodiment 1 lies in the length of the bark. Different bark lengths are set, as shown in the following Table 5.

[0088] Table 5. Different bark lengths

[0089] Group Bark length (cm) one 0.5 two 1.0 three 1.5 Four 2.0 five 2.5 six 3.0

[0090] The above six groups of different bark lengths were prepared into cultivation substrates according to Example 1 and cultivated with Anoectochilus roxburghii. Thirty well-developed Anoectochilus roxburghii seedlings were planted in each cultivation substrate. The average fresh weight growth rate (compared with the previous fresh weight) and the survival rate of each group of Anoectochilus roxburghii were calculated every 15 days. After 90 days, the proportion of the effective ingredients Anoectochilus roxburghii polysaccharide and Anoectochilus glycoside in the fresh weight of Anoectochilus roxburghii of different groups was detected. The fresh weight growth rate of Anoectochilus roxburghii is shown in Table 6 below, the mortality rate is shown in Table 7 below, the effective ingredient content is shown in Table 7 below, and the effective ingredient detection method is the same as the detection method of Anoectochilus roxburghii polysaccharide and Anoectochilus glycoside in Reference Example 2.

[0091] Table 6. Growth rate of fresh weight of golden thread vine planted in cultivation medium prepared with different bark lengths

[0092]

[0093]

[0094] Table 7. Survival rate of Anoectochilus roxburghii planted in cultivation medium prepared with different bark lengths

[0095] Group 15d 30d 45d 60d 75d 90d one 100％ 100％ 96.7％ 93.3％ 90％ 83.3％ two 100％ 96.7％ 96.7％ 93.3％ 90％ 86.7％ three 100％ 100％ 96.7％ 96.7％ 93.3％ 93.3％ Four 100％ 100％ 100％ 100％ 96.7％ 96.7％ five 96.7％ 96.7％ 93.3％ 93.3％ 90％ 90％ six 96.7％ 96.7％ 96.7％ 93.3％ 87.6％ 87.6％

[0096] Table 8. Content of active ingredients in the cultivation medium prepared with different bark lengths for planting Anoectochilus roxburghii

[0097] Active substance content Group 1 Group 2 Group 3 Group 4 Group 5 Group 6 Anoectochilus polysaccharide 18.1％ 19.4％ 24.7％ 33.5％ 18.3％ 17.6％ Anoectochilus glycoside 15.7％ 16.2％ 19.6％ 24.7％ 15.8％ 15.3％

[0098] According to the data analysis of Tables 6 and 7, the cultivation matrix prepared from the bark of Group 4 can make the fresh weight of Anoectochilus grow the most and the survival rate the highest within 90 days. According to the data analysis of Table 8, the cultivation matrix prepared from the bark of Group 4 makes the content of Anoectochilus polysaccharide and Anoectochilus glycoside in Anoectochilus significantly higher than that of the other groups. The bark contains rich humus after fermentation. Humus can not only provide nutrition for Anoectochilus, but also some substances contained in it can improve the disease resistance of Anoectochilus. The bark length of Groups 1 and 2 is too short, and the degradation rate of humus is too fast. Anoectochilus can absorb humus and grow fast in fresh weight in the early stage, but cannot absorb enough nutrients in the later stage, resulting in slow growth and low survival rate; the bark length of Groups 5 and 6 is too long, and humus is not easy to degrade, so Anoectochilus cannot absorb enough nutrients, resulting in slow growth and low survival rate; the bark length of Groups 3 and 4 is appropriate, which can enable Anoectochilus to absorb enough nutrients during the growth process. The bark length of Group 4 is more conducive to the growth of Anoectochilus and increase biomass and effective ingredients than Group 3.

[0099] So the bark length is preferably 2 cm.

[0100] Example 4: Effect of light intensity on the growth of Anoectochilus roxburghii

[0101] The difference between this embodiment and embodiment 1 is that the light intensity is different, and different light intensities are set, as shown in the following Table 9.

[0102] Table 9, different light intensities

[0103]

[0104]

[0105] The above seven groups of light intensity were planted and cultivated according to the cultivation method provided in Example 1. 30 well-developed golden thread vine seedlings were planted in each cultivation medium. The average fresh weight growth rate (compared with the previous fresh weight) and survival rate of each group of golden thread vine were calculated every 15 days. After 90 days, the proportion of the effective ingredients golden thread vine polysaccharide and golden thread glycoside in the fresh weight of golden thread vine in different groups was detected. The fresh weight growth rate of golden thread vine is shown in Table 10 below, the mortality rate is shown in Table 11 below, the effective ingredient content is shown in Table 12 below, and the effective ingredient detection method is the detection method of golden thread vine polysaccharide and golden thread glycoside in Reference Example 2.

[0106] Table 10. Growth rate of fresh weight of golden thread vine planted under different light intensities

[0107] Group 15d 30d 45d 60d 75d 90d one 15％ 28％ 38％ 46％ 63％ 71％ two 14％ 25％ 34％ 45％ 57％ 68％ three 14％ 23％ 33％ 42％ 54％ 59％ Four 12％ 20％ 28％ 35％ 48％ 55％ five 9％ 16％ 26％ 30％ 35％ 38％ six 7％ 13％ 17％ 21％ 26％ 31％ seven 5％ 9％ 13％ 17％ 21％ 24％

[0108] Table 11. Survival rate of golden thread vine planted at different light intensities

[0109] Group 15d 30d 45d 60d 75d 90d one 100％ 100％ 100％ 100％ 100％ 96.7％ two 100％ 100％ 100％ 100％ 96.7％ 96.7％ three 100％ 96.7％ 96.7％ 96.7％ 93.3％ 93.3％ Four 96.7％ 96.7％ 93.3％ 93.3％ 90％ 90％ five 90％ 83.3％ 76.6％ 70％ 66.7％ 63.3％ six 86.7％ 73.3％ 66.7％ 60％ 53.3％ 46.7％ seven 70％ 63.3％ 53.3％ 46.7％ 36.7％ 30％

[0110] Table 12. Content of active ingredients in Anoectochilus roxburghii planted under different light intensities

[0111] Active substance content Group 1 Group 2 Group 3 Group 4 Group 5 Group 6 Group 7 Anoectochilus polysaccharide 33.5％ 30.2％ 27.5％ 23.1％ 18.3％ 15.7％ 13.2％ Anoectochilus glycoside 24.7％ 21.6％ 19.3％ 17.2％ 15.8％ 13.5％ 11.6％

[0112] According to the data analysis of Tables 10 and 11, it is shown that the fresh weight growth rate and survival rate of the golden thread vine in groups 1 to 4 (light intensity less than 2000Lux) were higher than those in groups 5 to 7 (light intensity greater than 2000Lux) within 90 days of cultivation. When the light intensity was greater than 2000Lux, the fresh weight growth rate and survival rate of the golden thread vine decreased significantly with the increase of light intensity, so the light intensity for cultivating golden thread vine needs to be controlled within 2000Lux.

[0113] According to the data analysis in Table 12, the contents of Anoectochilus roxburghii polysaccharides and anoectochilus glycosides in Groups 1 to 4 (light intensity less than 2000 Lux) were higher than those in Groups 5 to 7 (light intensity greater than 2000 Lux) after 90 days, and the content of active ingredients in Anoectochilus roxburghii in Group 1 (light intensity of 500 Lux) was the highest compared with the other groups, indicating that a light intensity of 500 Lux is more conducive to the synthesis of active ingredients in Anoectochilus roxburghii.

[0114] The golden thread vine grown in the natural environment does not like light and prefers a cool environment, so its photosynthesis saturation point is low. Light intensity higher than 2000Lux will not only burn the golden thread vine and cause its survival rate to decrease, but also affect its photosynthesis efficiency, resulting in a decrease in the content of synthetic effective ingredients.

[0115] Preferably, the light intensity is 500 Lux.

[0116] Example 5: Effects of different light treatment and dark treatment durations on the synthesis of active ingredients in Anoectochilus roxburghii

[0117] The difference between this embodiment and embodiment 1 is that the light treatment duration and the dark treatment duration are different, as shown in Table 13 below.

[0118] Table 13. Different light treatment and dark treatment durations

[0119] Group Light treatment time (h) Dark processing time (h) one 18 6 two 16 8 three 14 10 Four 12 12 five 10 14 six 8 16

[0120] The above six groups of different light treatment and dark treatment durations were planted and cultivated according to the cultivation method provided in Example 1. Thirty well-developed golden thread seedlings were planted in each group. After 90 days, the proportion of the effective ingredients golden thread polysaccharide and golden thread glycoside in the fresh weight of golden thread in different groups was detected. The content of the effective ingredients is shown in Table 14 below. The detection method of the effective ingredients refers to the detection method of golden thread polysaccharide and golden thread glycoside in Example 2.

[0121] Table 14. Content of active ingredients in Anoectochilus roxburghii planted under different light treatment and dark treatment durations

[0122] Active substance content Group 1 Group 2 Group 3 Group 4 Group 5 Group 6 Anoectochilus polysaccharide 33.5％ 28.4％ 25.3％ 20.7％ 14.2％ 11.9％ Anoectochilus glycoside 24.7％ 22.1％ 18.6％ 15.5％ 12.6％ 9.0％

[0123] According to the data analysis in Table 14, it was found that the dark treatment time of Group 1 to Group 6 gradually increased, and the contents of Anoectochilus roxburghii polysaccharides and Anoectochilus glycosides in Anoectochilus roxburghii gradually decreased from Group 1 to Group 6. Therefore, the contents of Anoectochilus roxburghii polysaccharides and Anoectochilus glycosides in Anoectochilus roxburghii were inversely proportional to the dark treatment time. Too long dark treatment time will cause the content of effective ingredients consumed by respiration to be greater than the content of effective ingredients synthesized by photosynthesis, resulting in a decrease in the content of effective ingredients in Anoectochilus roxburghii.

[0124] Preferably, the light treatment time is 18 hours and the dark treatment time is 6 hours in one day.

[0125] Although the present invention is disclosed as above, the present invention is not limited thereto. Any person skilled in the art can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the scope defined by the claims.

Claims

1. A culture medium for Anoectochilus roxburghii, characterized in that: Including gravel, granite, perlite, bark, river sand, and peat soil.

2. The anoectochilus roxburghii cultivation medium according to claim 1, characterized in that The mixing mass ratio of the gravel, granite, perlite, bark, river sand and peat soil is (2.4-3.6): (2.4-3.6): (0.8-1.2): (1.6-2.4): (0.8-1.2): (8-12)。 3. The anoectochilus roxburghii cultivation medium according to claim 2, characterized in that The diameters of the gravel and granite are 1-5 cm, the diameter of the perlite is 0.3-3 mm, the length of the bark is 1-3 cm, the diameter of the river sand is 0-0.5 mm, and the thickness of the peat soil is 20-40 mm.

4. The anoectochilus roxburghii cultivation medium according to claim 3, characterized in that: The length of the bark is preferably 2 cm.

5. A method for cultivating golden lotus, characterized in that: Using the anoectochilus roxburghii cultivation substrate according to claims 1-4, the cultivation method comprises the following steps: (1) Sterilizing the cultivation medium; (2) Transplanting the roxburghii seedlings into the cultivation medium and cultivating them in an indoor environment; (3) Adjust the humidity of the cultivation medium and the humidity and temperature of the indoor environment, as well as the duration, brightness, and illumination method of the lighting.

6. The method for cultivating golden lotus according to claim 5, characterized in that: The sterilization method is 121° C., high pressure sterilization for 15-20 minutes.

7. The method for cultivating golden lotus according to claim 5, characterized in that: The humidity of the cultivation medium is maintained at 40%-50%, the air humidity is maintained at 65%-70%, the air humidity is maintained at 65%-70%.

8. The method for cultivating golden lotus according to claim 5, characterized in that: Within a day, the illumination duration is 18 hours, the light brightness is maintained within 2000Lux, the illumination mode is oblique or diffuse reflection, and the temperature is controlled at 29℃-30℃; the no-light duration is 6 hours, and the temperature is controlled at 18℃-23℃.

9. A use of a matrix composition for increasing the biomass of Anoectochilus roxburghii, characterized in that: The matrix composition includes gravel, granite, perlite, bark, river sand, and peat soil.

10. A use of a matrix composition for improving the growth and accumulation of roxburghii polysaccharides and roxburghii glycosides in Lycopodiella cinerea, characterized in that: The matrix composition includes gravel, granite, perlite, bark, river sand, and peat soil.

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