Ecological cultivation method of gentiana veitchiorum and preparation and application of gentiana veitchiorum extract

Abstract

The invention relates to a method for promoting the rooting of gentiana blue, which adopts glacier water to treat gentiana blue seeds or cuttings, promotes the germination of gentiana blue seeds or the rooting of cuttings, improves the germination rate, rooting rate, rooting speed and rooting time of gentiana blue, and also provides an ecological cultivation method of gentiana blue. The method has the advantages that seedling raising and large-scale propagation are carried out on the gentiana veitchiorum, successful transplanting and planting are achieved, the cost is low, land and labor are saved, the environment is protected, popularization is easy, and economic benefits are high, a good foundation is laid for manual large-scale planting of the gentiana veitchiorum, and good economic benefits, social benefits and ecological benefits are achieved. The gentiana veitchiorum flowers obtained through ecological cultivation are extracted with different solvents, the obtained gentiana veitchiorum extract is rich in active ingredients and stable in quality, and the prepared extract has the effects of oxidation resistance, aging resistance, inflammation resistance, whitening and the like and has good application prospects in the industries of cosmetics, health care products and pharmacy.

Description

Technical Field

[0001] The invention relates to the field of plant cultivation, and in particular to an ecological cultivation method of blue hosta gentiana and preparation and application of an extract thereof. Background Art

[0002] Gentiana is one of the traditional Chinese medicinal materials. Its main functions are to clear away heat and dampness, and to clear liver fire. It is used to treat damp-heat jaundice, cholecystitis, loss of appetite, red eyes, urinary tract infection, herpes zoster and hyperthyroidism. It is an important raw material for nearly 200 Chinese patent medicines. Among them, the prescription "Longdan Xiegan Decoction" has made Gentiana famous throughout the ages. Blue Hosta Gentiana is the most widely used plant in Tibetan medicine. In Tibetan medicine, its flowers are used to treat smallpox, tracheitis and cough, and the whole plant is used to treat red eyes and headaches, pharyngitis and damp-heat jaundice. It is also often used as a medicine in compound prescriptions in Tibetan medicine.

[0003] Most of the Gentiana plants in Tibet grow in high-altitude mountains. The harsh natural environment forces them to accumulate more dry matter and synthesize more natural ingredients during their growth, so as to help the plants adapt to the plateau environment. However, the Gentiana plants in Tibet are basically in the wild state at present. Therefore, exploring and studying their reproduction and cultivation techniques is of great significance to the effective protection and scientific utilization of the Gentiana plant resources in Tibet.

[0004] The gentian currently grown on a large scale in the market is Yunnan Gentiana yunnanensis ( GentianarigescensFranch .ex Hemsl.), Gentiana from the main producing areas of Northeast China ( GentianascabraBunge ), and the main production in Europe is Gentiana lutea ( Gentianalutea L.) and other varieties. Different species of gentian have different growth environments and plant characteristics, which result in completely different breeding conditions and methods.

[0005] Blue Hosta Gentiana Gentianaveitchiorum Hemsl.), its seeds are small and the seedlings grow slowly, so seed propagation and seedling preservation are somewhat difficult in production. In addition, the blue hosta gentian grows in specific areas of the plateau, with high altitudes, thin air, and large temperature differences between day and night. It has special requirements for soil, water, and sunlight, and it is difficult to propagate using conventional methods.

[0006] The blue hosta gentian currently on the market is all wild, and the quality of the medicinal materials varies. However, due to its high medicinal value, it has been harvested in large quantities, and the market is in short supply. The large-scale harvesting and use of wild gentian will accelerate the depletion of Tibetan medicinal resources and gradually reduce wild resources, which greatly limits the development and utilization of blue hosta gentian resources, a Chinese characteristic medicinal plant. Summary of the invention

[0007] The invention uses the seed propagation and cutting propagation technologies to carry out ecological seedling raising and large-scale propagation of the blue hosta gentian, and successfully transplants and plants it.

[0008] One aspect of the present invention provides a method for promoting the germination of blue hosta gentiana seeds and / or the rooting of cuttings. Wherein, the method comprises the following steps: 1) Pre-treating the seeds and / or cuttings of Gentiana scabra; The pre-treatment is to use glacier water and / or deionized water to treat seeds and / or cuttings, and the treatment time is 20 to 72 hours; 2) Planting the treated blue hosta gentian seeds and / or cuttings into a seedling medium and / or a cutting medium, and controlling the temperature and humidity of the growth environment, thereby promoting the germination of the blue hosta gentian seeds and / or the rooting of the cuttings; The seed germination temperature is 20-30°C, the soil humidity is above 30%, and the air humidity is 60%-70%; The rooting temperature of the cuttings is 18-28° C., and the humidity is above 85%.

[0009] Furthermore, before using the seedling medium and / or cutting medium, it is steam sterilized at 120° C. for 30 minutes and used after cooling.

[0010] Preferably, when promoting the germination of blue hosta gentian seeds, the following steps are performed: using glacial water and / or deionized water to soak the seeds for 20 to 72 hours, adding the seeds and the seedling medium after the rooting promotion treatment into a container with vermiculite at the bottom, and then covering the top with a layer of vermiculite to promote rooting; Wherein, the seedling culture medium comprises fine sand or fine sawdust.

[0011] Furthermore, when the seeds are stored for a long time, the blue hosta gentian seeds are germinated to promote seed germination, and the germination includes the following steps: i) Soak the seeds in 0.05-0.2 ml / L of gibberellin in glacial water and / or deionized water for 20-72 hours, remove and rinse with clean water. ii) Mix the seeds and fine sand evenly, put them into a container, place it in a sunny place, cover the container with wet gauze for germination, stabilize the temperature at 22-25℃, and ensure that the fine sand has a certain temperature, and cultivate for 5-7 days.

[0012] The glacier water is collected from the Qudennima Spring in Tibet, Himalayas. The Dorje Qudennima Spring is located on the northern slope of the Himalayas, 5,128 meters above sea level, and the temperature of the spring mouth is maintained at 1.5-2°C all year round. Further, when promoting the rooting of blue hosta gentian cuttings, the following steps are included; Prepare a rooting solution with an IBA content of 5 to 150 mg / kg by mixing glacier water and / or deionized water with IBA, and soak the blue hosta gentian cuttings in the prepared rooting solution for 20 to 72 hours; Inserting the pretreated blue hosta gentian cuttings into a cutting medium at a temperature of 20 to 30° C., a soil humidity of more than 30%, and an air humidity of 60% to 70% to promote rooting of the blue hosta gentian cuttings, wherein the cutting medium contains vermiculite; The glacier water is collected from the Qudennima Spring in Tibet, the Himalayas. The Doji Qudennima Spring is located on the northern slope of the Himalayas, 5,128 meters above sea level. The temperature at the spring mouth remains at 1.5 to 2°C all year round.

[0013] Furthermore, the blue hosta gentiana cuttings rooting treatment also includes the following steps: i) Seedling strengthening: Collect blue hosta gentian seedlings and transplant them to the greenhouse for seedling strengthening. The temperature is controlled at 18-28℃ and the humidity is controlled at more than 85%. Plant for more than 10 days; ii) Cut the cuttings from the blue hosta gentian seedlings, select strong and disease-free tender branches, cut them into 2-3 cm cuttings at the lower part of the branches, make the cuts smooth, remove the base leaves, and trim the upper leaves of the cuttings appropriately; iii) Sterilize the base of the cuttings with 0.03% KMnO4 for 5 to 10 minutes.

[0014] Another aspect of the present invention provides an ecological cultivation method of blue hosta gentian, comprising the following steps: 1) using the above-mentioned rooting promotion method to promote the germination of blue hosta gentian seeds and / or the rooting of cuttings to obtain rooted blue hosta gentian seedlings; 2) Seedling cultivation, the seedling cultivation process includes the following steps: Transplant the rooted blue hosta gentian seedlings into the seedling tray containing the culture medium. The rooted seedlings are sprayed with fertilizer, the fertilizer is "Pendefu", the application concentration is 400 to 1200 times, preferably, the application concentration is 600 times, and the vermiculite surface is sterilized with a 0.03% KMnO4 solution every half a month after the seedlings emerge. The seedling cultivation conditions are room temperature of 18-28°C, humidity of more than 85%, shade, and watering 2-3 times a week; The cultivation matrix is ​​a mass ratio of perlite: garden soil = 0.5-1.5: 1.5-2.5, the soil binder is HPMC, and the concentration of the HPMC is 200-400 mg / L. Further, the perlite: garden soil = 1: 2, the concentration of the HPMC is 300 mg / L, the pH of the culture matrix is ​​5.7 ~ 6.79;

[0015] 3) Large-scale transplanting Gentiana seedlings are transplanted to the field after one year of growth. The transplanting time is in early to mid-April in spring or late September to early October in autumn. Before transplanting, the seedlings are placed outside the greenhouse for 7 to 10 days of hardening. The transplanting method is to cover the ground with a film, drill holes for planting, plant 2 to 3 gentian seedlings in each hole, and plant them at a spacing greater than 20 cm. During transplanting, continuous sprinkling irrigation is performed for at least 3 days. Before flowering, in mid-to-late July, organic fertilizer is applied, wherein the organic fertilizer is a fertilizer containing a humic acid aqueous solution.

[0016] Another aspect of the present invention provides a method for preparing an extract of Gentiana cyanus, comprising the following steps: 1. Collect blooming blue hosta gentiana flowers, dry them at 35-45°C, crush them and sieve them to obtain blue hosta gentiana flower powder; 2. Take the blue hosta gentian flower powder, add the solvent at a solid-liquid ratio of 1:15-100, perform ultrasonic extraction at 20-50°C with an ultrasonic power of 400-700W for more than 30 minutes, and then centrifuge and filter at 10000rpm to obtain the blue hosta gentian flower extract; Further, the solvent is one or more of glacier water, deionized water, ethanol, and butanediol. Further, the solvent is deionized water and / or glacier water. The material-liquid ratio is preferably 1:20, 1:50, and the ultrasonic power is 600W.

[0017] Another aspect of the present invention provides a skin external preparation with the blue hosta gentiana flower extract as an active ingredient and having anti-aging, whitening, anti-inflammatory, soothing and antioxidant functions.

[0018] Furthermore, a skin topical preparation is provided, which has a blue hosta gentian flower extract as an active ingredient, and has the functions of scavenging free radicals, inhibiting tyrosinase activity, inhibiting melanin production, preventing oxidative damage, and inhibiting TNF-α, IL-1α, IL-6 and / or IL-8 gene expression. The present invention uses seed propagation and cutting propagation techniques to carry out ecological seedling cultivation and large-scale propagation of blue hosta gentian, and successfully transplants and plants it.

[0019] One aspect of the present invention provides a method for promoting the germination of blue hosta gentiana seeds and / or the rooting of cuttings. Wherein, the method comprises the following steps: 1) Pre-treating the seeds and / or cuttings of Gentiana scabra; The pre-treatment is to use glacier water and / or deionized water to treat seeds and / or cuttings, and the treatment time is 20 to 72 hours; 2) Planting the treated blue hosta gentian seeds and / or cuttings into a seedling medium and / or a cutting medium, and controlling the temperature and humidity of the growth environment, thereby promoting the germination of the blue hosta gentian seeds and / or the rooting of the cuttings; The seed germination temperature is 20-30°C, the soil humidity is above 30%, and the air humidity is 60%-70%; The rooting temperature of the cuttings is 18-28° C., and the humidity is above 85%.

[0020] Furthermore, before using the seedling medium and / or cutting medium, it is steam sterilized at 120° C. for 30 minutes and used after cooling.

[0021] Preferably, when promoting the germination of blue hosta gentian seeds, the following steps are performed: using glacial water and / or deionized water to soak the seeds for 20 to 72 hours, adding the seeds and the seedling medium after the rooting promotion treatment into a container with vermiculite at the bottom, and then covering the top with a layer of vermiculite to promote rooting; Wherein, the seedling culture medium comprises fine sand or fine sawdust.

[0022] Furthermore, when the seeds are stored for a long time, the blue hosta gentian seeds are germinated to promote seed germination, and the germination includes the following steps: i) Soak the seeds in 0.05-0.2 ml / L of gibberellin in glacial water and / or deionized water for 20-72 hours, remove and rinse with clean water. ii) Mix the seeds and fine sand evenly, put them into a container, place it in a sunny place, cover the container with wet gauze for germination, stabilize the temperature at 22-25℃, and ensure that the fine sand has a certain temperature, and cultivate for 5-7 days.

[0023] The glacier water is collected from the Qudennima Spring in Tibet, Himalayas. The Dorje Qudennima Spring is located on the northern slope of the Himalayas, 5,128 meters above sea level, and the temperature of the spring mouth is maintained at 1.5-2°C all year round. Further, when promoting the rooting of blue hosta gentian cuttings, the following steps are included; Prepare a rooting solution with an IBA content of 5 to 150 mg / kg by mixing glacier water and / or deionized water with IBA, and soak the blue hosta gentian cuttings in the prepared rooting solution for 20 to 72 hours; Inserting the pretreated blue hosta gentian cuttings into a cutting medium at a temperature of 20 to 30° C., a soil humidity of more than 30%, and an air humidity of 60% to 70% to promote rooting of the blue hosta gentian cuttings, wherein the cutting medium contains vermiculite; The glacier water is collected from the Qudennima Spring in Tibet, the Himalayas. The Doji Qudennima Spring is located on the northern slope of the Himalayas, 5,128 meters above sea level. The temperature at the spring mouth remains at 1.5 to 2°C all year round.

[0024] Furthermore, the blue hosta gentiana cuttings rooting treatment also includes the following steps: i) Seedling strengthening: Collect blue hosta gentian seedlings and transplant them to the greenhouse for seedling strengthening. The temperature is controlled at 18-28℃ and the humidity is controlled at more than 85%. Plant for more than 10 days; ii) Cut the cuttings from the blue hosta gentian seedlings, select strong and disease-free tender branches, cut them into 2-3 cm cuttings at the lower part of the branches, make the cuts smooth, remove the base leaves, and trim the upper leaves of the cuttings appropriately; iii) Sterilize the base of the cuttings with 0.03% KMnO4 for 5 to 10 minutes.

[0025] Another aspect of the present invention provides an ecological cultivation method of blue hosta gentian, comprising the following steps: 1) using the above-mentioned rooting promotion method to promote the germination of blue hosta gentian seeds and / or the rooting of cuttings to obtain rooted blue hosta gentian seedlings; 2) Seedling cultivation, the seedling cultivation process includes the following steps: Transplant the rooted blue hosta gentian seedlings into the seedling tray containing the culture medium. The rooted seedlings are sprayed with fertilizer, the fertilizer is "Pendefu", the application concentration is 400 to 1200 times, preferably, the application concentration is 600 times, and the vermiculite surface is sterilized with a 0.03% KMnO4 solution every half a month after the seedlings emerge. The seedling cultivation conditions are room temperature of 18-28°C, humidity of more than 85%, shade, and watering 2-3 times a week; The cultivation matrix is ​​a mass ratio of perlite: garden soil = 0.5-1.5: 1.5-2.5, the soil binder is HPMC, and the concentration of the HPMC is 200-400 mg / L. Furthermore, the ratio of perlite to garden soil is 1:2, the concentration of HPMC is 300 mg / L, and the pH of the culture matrix is ​​5.7-6.79.

[0026] 3) Large-scale transplanting Gentiana seedlings are transplanted to the field after one year of growth. The transplanting time is in early to mid-April in spring or late September to early October in autumn. Before transplanting, the seedlings are placed outside the greenhouse for 7 to 10 days of hardening. The transplanting method is to cover the ground with a film, drill holes for planting, plant 2 to 3 gentian seedlings in each hole, and plant them at a spacing greater than 20 cm. During transplanting, continuous sprinkling irrigation is performed for at least 3 days. Before flowering, in mid-to-late July, organic fertilizer is applied, wherein the organic fertilizer is a fertilizer containing a humic acid aqueous solution.

[0027] Another aspect of the present invention provides a method for preparing an extract of Gentiana cyanus, comprising the following steps: 1. Collect blooming blue hosta gentiana flowers, dry them at 35-45°C, crush them and sieve them to obtain blue hosta gentiana flower powder; 2. Take the blue hosta gentian flower powder, add the solvent at a solid-liquid ratio of 1:15-100, perform ultrasonic extraction at 20-50°C with an ultrasonic power of 400-700W for more than 30 minutes, and then centrifuge and filter at 10000rpm to obtain the blue hosta gentian flower extract; Further, the solvent is one or more of glacier water, deionized water, ethanol, and butanediol. Further, the solvent is deionized water and / or glacier water. The material-liquid ratio is preferably 1:20, 1:50, and the ultrasonic power is 600W.

[0028] Another aspect of the present invention provides a skin external preparation with the blue hosta gentiana flower extract as an active ingredient and having anti-aging, whitening, anti-inflammatory, soothing and antioxidant functions.

[0029] Furthermore, a skin topical preparation is provided which contains a blue hosta gentian flower extract as an active ingredient and has the functions of scavenging free radicals, inhibiting tyrosinase activity, inhibiting melanin production, preventing oxidative damage, and inhibiting TNF-α, IL-1α, IL-6 and / or IL-8 gene expression.

[0030] Beneficial Effects 1) The ecological cultivation method of the blue hosta gentian of the present invention, through raising seedlings and large-scale propagation of the blue hosta gentian and successfully transplanting and planting, has the advantages of simple operation, low cost, saving land and labor, protecting the environment, easy promotion, and high economic benefit, laying a good foundation for the artificial large-scale planting of the blue hosta gentian, and having good economic benefits, social benefits and ecological benefits.

[0031] 2) The present invention pre-treats the seeds or cuttings of the blue hosta gentian, promotes the germination of the blue hosta gentian seeds or the rooting of the cuttings, improves the rooting rate, rooting speed and rooting time, and greatly improves the survival rate of the blue hosta gentian.

[0032] 3) The present invention also extracts the flowers of the blue hosta gentian with different solvents, and the obtained blue hosta gentian extract has a stable source of raw materials. After detection and analysis, the extract has rich active ingredients and stable quality between batches. The prepared extract also has antioxidant, anti-aging, anti-inflammatory, whitening and other effects, and has a good application prospect in the cosmetics, health care products and pharmaceutical industries. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 Example 2 Comparison of the planting effects of pure water treatment of blue hosta gentian cuttings (a) and glacier water + IBA (b) Figure 2 Example 3 Transplantation of blue hosta gentian seedlings Figure 3 Blue Hosta Gentiana plantation (a), picked blue Hosta Gentiana flowers (b) Figure 4 Comparison of blue hosta gentiana flower extracts obtained by different extraction methods in zebrafish whitening test Implementation

[0034] The present invention is described in more detail below with reference to examples, but these are not intended to limit the technical scope of the present invention.

[0035] The raw materials used in the embodiments of the present invention are as follows: Perlite specifications: particle size 4-7mm; vermiculite particle size: 3-5mm, and garden soil are commonly used gardening materials.

[0036] Glacier water: Glacier water is collected from Qudennima in the Himalayas of Tibet. The local Dorje Qudennima Spring is located on the northern slope of the Himalayas, 5,128 meters above sea level. The temperature at the spring mouth remains at 1.5-2°C all year round.

[0037] IBA: indolebutyric acid Hefei BASF Biotechnology Co., Ltd.; Gibberellic acid: Shenzhen Love Biotechnology Co., Ltd.; Potassium permanganate: Guangzhou Jinzhisheng Chemical Co., Ltd.; Pendefu: Pendefu foliar fertilizer Shanghai Yongtong Ecological Engineering Co., Ltd.; HPMC: Hydroxypropyl methylcellulose, viscosity about 200,000, Guizhou Fuqiang Fine Chemical Co., Ltd.; Fertilizer containing humic acid aqueous solution: Beiling humic acid water-soluble fertilizer Beijing Jinggeng Tianxia Agricultural Technology Co., Ltd.

[0038] Example 1 Promoting the germination of blue hosta gentiana seeds

[0039] 1) Before sowing, take 400 seeds of Gentiana cyanobacteria and divide them into two groups. Soak the seeds in 0.1 ml / L gibberellin in glacial water and / or deionized water for 24 h. 2) Mix seeds and fine sand in a mass ratio of 1:4, put them into a container, place it in a sunny place indoors, cover the container with wet gauze for germination, stabilize the temperature at 22-25℃, and ensure that the fine sand has a certain temperature. Cultivate for 6 days and sow when the seeds turn white.

[0040] Divide 200 seeds into two groups, mix them with 15 times the weight of fine sand and 170 times the weight of fine sawdust, stir well and sprinkle evenly into a fully moistened vermiculite basin. After sowing, cover with a layer of vermiculite to keep moisture. Control the temperature at 20-30℃, soil moisture above 30%, and air humidity at 60%-70% to promote its germination.

[0041] The average number of days for seeds to germinate after passing through glacial water and deionized water was counted, and the number of germinations is shown in Table 1: Table 1 Statistics of germination rate and germination time of Gentiana scabra seeds

[0042] When seeds germinate, they usually grow roots first and then sprouts. By counting the germination time and number of seeds, the effects of promoting rooting in two different ways were evaluated. As shown in Table 1, glacial water can shorten the number of seed germination days, increase the number of blue hosta gentian seeds germination, shorten the germination time, and effectively promote seed germination. Moreover, the number of germination in the fine sand substrate is better than that in fine sawdust, and the germination time of fine sawdust is shorter than that of fine sand.

[0043] Collect blue hosta gentian seedlings and transplant them into the greenhouse for seedling growth. The temperature is controlled at 18-28℃ and the humidity is controlled at above 85%. After 10 days of adaptation, cut branches suitable for cuttings from the plants for cutting rooting to obtain seedlings. Select tender branches with consistent growth, strong growth, and no diseases and insect pests as cuttings, cut them at the bottom of the branches, cut them into 2-3cm cuttings, make the cuts smooth, remove the base leaves, and properly trim the upper leaves of the cuttings. This can effectively inhibit the transpiration of the cuttings, weaken the respiration, reduce the consumption of nutrients in the cuttings, increase the accumulation of organic matter and auxin, and promote the rapid rooting of the cuttings.

[0044] The collected cuttings are sterilized with 0.03% KMnO4 for 5 to 10 minutes. The characteristic of Gentiana scabra to take root at the branches is fully utilized during cuttings, so that it can take root at the branches and absorb nutrients and water in the soil for the survival and growth of the cuttings.

[0045] The cuttings were treated with pure water, glacier water, and 10 mg / Kg IBA prepared with pure water and glacier water. 180 cuttings were treated in each group and repeated in 3 groups. The cuttings were inserted into the matrix filled with vermiculite. The cutting indicators were recorded after 60 days, including rooting rate, average number of roots, and rooting time. The planting effect of blue hosta gentian cuttings treated with pure water (a) and glacier water + IBA (b) is shown in Figure 2 The test results are as follows: Table 2 shows the effects of different treatments on the rooting rate of blue hosta gentian cuttings. It can be seen from Table 2 that the four different treatments can make blue hosta gentian take root, and the use of glacier water and 10 mg / Kg IBA can significantly improve the rooting rate. The rooting rates of the four groups were 17.78, 24.93, 34.44% and 46.76%, respectively.

[0046] Table 2 Effects of different treatments on the rooting rate of Gentiana scabra cuttings project Pure water Glacier water Pure water + IBA Glacier water + IBA Number of cuttings 30 30 30 30 Rooting rate of the first group (%) 16.67 25.38 36.67 40.32 Rooting rate of the second group (%) 23.33 28.62 33.33 48.31 Rooting rate of the third group (%) 13.33 20.78 33.33 45.21 Average rooting rate (%) 17.78 24.93 34.44 46.76 Note: Rooting rate = (number of rooted cuttings / number of tested cuttings) × 100% Table 3 Effects of different treatments on the average rooting number of cuttings of Hosta gentiana project Pure water Glacier water Pure water + IBA Glacier water + IBA Number of cuttings 30 30 30 30 Average number of roots in the first group (pcs) 2.2 2.8 3.41 4.41 Average number of roots in the second group (pcs) 1.29 1.89 4.7 4.90 Average number of roots in the third group 2 2.48 3 3.28 Average number of roots in three groups 1.83 2.39 3.7 4.20 Note: Average rooting number = total number of rooted cuttings / total number of rooted cuttings As shown in Table 3, different treatments have different effects on the number of roots of Hosta gentiana. The use of glacier water and glacier water + 10 mg / Kg IBA can increase the number of roots. The number of roots in the four treatments are 1.83, 2.39, 3.7 and 4.20, respectively.

[0047] Table 4 shows the effects of different treatments on the rooting time of blue hosta gentian cuttings project Pure water Glacier water Pure water + IBA Glacier water + IBA Number of cuttings 30 30 30 30 Rooting time (days) 18 16 16 14 As shown in Table 4, different treatment methods have different effects on the rooting time of Hosta gentiana. The rooting time of the pure water treatment group is the longest, 18 days. Glacier water and IBA can shorten the rooting days, both of which are 16 days. The rooting days of glacier water mixed with IBA are 14 days.

[0048] Table 5 Effects of different treatments on the rooting time of terminal buds of Hosta gentiana Table 5 shows the effects of different treatments on the rooting time of the terminal buds of Blue Hosta Gentiana. It can be seen from Table 5 that although the different treatments have a certain impact on the rooting days of Blue Hosta Gentiana, the rooting time is 10-12 days. The rooting days of the terminal bud cuttings of glacier water + IBA are shorter. Using its terminal bud rooting can shorten its rooting days.

[0049] In summary, using glacier water to treat the cuttings of blue hosta gentian can promote its rooting, and using glacier water instead of pure water to prepare IBA can also achieve the effect of promoting rooting. Using glacier water to propagate blue hosta gentian is green, environmentally friendly and low-cost. In addition, using the terminal buds of blue hosta gentian for rooting can shorten the rooting time of gentian and quickly obtain gentian seedlings for propagation.

[0050] In addition, after the blue hosta gentian is grafted, the new roots are tender and brittle compared to the roots of the wild blue hosta gentian, and the degree of lignification is relatively low. It should be transplanted after the lateral roots are differentiated to form a relatively complete root system and the root system has a high degree of lignification. In the early stage of transplanting, care should be taken to avoid strong sunlight, and shade should be provided. After the plants adapt to the transplanting environment, the shade cloth can be removed.

[0051] Example 3 Ecological cultivation of Gentiana cyanea 1) Methods to promote rooting The method of Example 1 and Example 2 was used to treat the seeds / cuttings to obtain the blue hosta gentian seedlings; 2) Cultivation of blue hosta seedlings Transplant the gentian seedlings into a seedling tray containing a culture medium; prepare the culture medium with perlite and garden soil at a ratio of 1:2 and HPMC 300mg / L as soil binder. The pH of the culture medium is 5.7-6.79. The soil compactness is closest to the soil environment where the blue hosta grows under this concentration and matrix combination. Under this soil quality, transplanting the seedlings with the soil ball can improve the survival rate of the gentian seedlings.

[0052] Spraying the rooted blue hosta gentian seedlings with 600 times the "Pendefu" fertilizer can effectively increase the number and length of leaves and the height of the plant. At the same time, 0.25% to 0.5% calcium nitrate can be sprayed. When calcium nitrate is used as a topdressing, because the nutrients of calcium nitrate fertilizer are easy to be lost, it should be used in small amounts and multiple times to avoid unnecessary fertilizer waste, so as to improve the toughness of the gentian rhizomes.

[0053] After the seedlings emerge, the surface of the vermiculite is sterilized with a 0.03% KMnO4 solution every half a month. The seedlings are cultured at room temperature of 18-28°C, humidity of more than 85%, shaded, and watered 2-3 times a week for moisture retention. Therefore, each watering should be done until the surface is 2-3 cm wet to keep the leaves moist.

[0054] 3) Large-scale transplanting After one year of growth, the cutting seedlings are transplanted to the field. The transplanting time is in early April in spring. The seedlings need to be placed outside the greenhouse for 7 days before transplanting. The mulch-covered hole-seeding cultivation technology is adopted. On the basis of mulch-covered, holes are drilled with an eye-drilling device for hole-seeding. 2 to 3 seedlings are planted in each hole. This not only increases the temperature of the soil surface, but also plays a role in collecting rainwater and conserving moisture, and effectively increases the water content of the cultivated soil. At the same time, hole-seeding increases the uniformity of planting, inhibits the growth of weeds, and thus improves the efficiency of field management, achieving the purpose of saving labor, increasing production and increasing income. Weeding should be carried out at any time during the growth period. When pulling out weeds close to the seedlings, be sure to press the soil around the seedlings to prevent affecting the roots of the seedlings and ensure the normal growth of the seedlings. In mid-July, dig trenches between rows and apply 500 times of Belin humic acid aqueous solution fertilizer (humic acid ≥ 30g / L, N+P2O5+K2O ≥ 200g / L, 20Kg / barrel) to increase the number of flower buds and fruit setting rate, promote seed maturity, and make the grains plump. Figure 3 a.

[0055] Example 4 Preparation of Blue Hosta Gentiana Flower Extract

[0056] In order to reduce the damage to the blue hosta gentian plants, the flowers of the blue hosta gentian obtained from ecological planting are developed. From September to October every year, the blooming blue hosta gentian flowers are collected. Figure 3 b. Dry at 40℃, sieve through a 40-mesh sieve to obtain blue hosta gentian flower powder. Take 10g of blue hosta gentian flower powder, add pure water and glacial water at a ratio of 1:50 for extraction, the extraction temperature is 35℃, 600W ultrasound for 30 min, centrifuge at 10000rpm and filter, take the filtrate to test its solid content between 0.8-1%, and the obtained extract is tested for efficacy.

[0057] 4.1 Component Analysis (1) Determination of total flavonoids Standard solution: Use quercetin to prepare an ethanol solution of 0.2315 mg / mL, and dilute it into a concentration gradient of 2-10 times. Take 300μL of the extract of long-leaf snow lotus callus, first add 50μL 5% NaNO2, mix, and place at room temperature for 6 min; then add 50μL 10% Al(NO3)3, mix, and place at room temperature for 6 min; finally add 500μL 4% NaOH and 140μL water, mix, and place at room temperature for 15 min. Take 100μL of the mixed solution to measure the OD value at 500 nm. The total polyphenol content of blue hosta gentian flower was calculated according to the standard curve. The results are shown in Table 6-Table 7: The average total flavonoid content of pure water and glacier water of blue hosta gentian flower is 1.0 mg / mL and 0.9 mg / mL, respectively.

[0058] (2) Polysaccharide determination The polysaccharide content was detected by the phenol-sulfuric acid method: 2 mL of the sample was added with 1 mL of 5% phenol reagent, and then 5 mL of concentrated sulfuric acid, and then allowed to stand for 5 min, and then heated in a 40°C water bath for 30-40 min, taken out, cooled to room temperature, and finally tested the absorbance at 490 nm, and the polysaccharide content was calculated according to the glucose standard solution curve. The results are shown in Tables 6 and 7: The average polysaccharide content of pure water, glacier water, and 10% of the blue hosta gentian flower was 4.3 mg / mL and 3.9 mg / mL, respectively, and the polysaccharide content obtained by extraction was the highest.

[0059] (3) Determination of total iridoid glycosides The standard curve was prepared by spectrophotometry: 5.68 mg of gentiopicroside reference substance was added with 25 mL of methanol to make a 0.227 mg / mL reference substance solution, which was diluted to 0.1135, 0.05675, 0.02837, 0.01418, 0.007093, and 0 mg / mL concentration gradients in sequence. 200 uL of standard substance and sample were taken to measure their absorbance at 270 nm. The total iridoid glycoside content was calculated based on the standard solution curve of gentiopicroside. The results are shown in Tables 6 and 7. The average contents of total iridoid glycosides in pure water, glacier water, and blue hosta gentiana were 4.2 mg / mL and 4.6 mg / mL, respectively.

[0060] (4) Determination of total polyphenols Standard curve: 0.33 mg / mL gallic acid solution, diluted to 0, 0.0052, 0.0103, 0.0206, 0.0413, 0.0825, 0.16500.3300 mg / mL concentration gradient. Take 100 μL of blue hosta gentian flower extract, add 500 μL 10% Folin-Ciocalteu reagent, mix thoroughly, place for 5 min (not more than 8 min), then add 400 μL 7.5% Na2CO3, mix thoroughly, room temperature for 60 min, and finally take 100 μL μL to measure the 765nm OD value on the microplate reader. The total polyphenol content of blue hosta gentian flower was calculated according to the standard curve, and the results are shown in Table 5-Table 7: The average content of total polyphenols in pure water and glacier water extracts of blue hosta gentian flower were 0.6 mg / mL and 0.6 mg / mL, respectively.

[0061] Table 6 Active ingredients of pure water extract of blue hosta gentiana flower Blue Hosta Gentiana Flower (Pure Water Extract) Total flavonoids mg / mL Polysaccharide mg / mL Total iridoid glycosides mg / mL Total polyphenols mg / mL Batch 1 1.0 4.3 4.3 0.6 Batch 2 0.9 4.2 4.1 0.5 Batch 3 1.0 4.5 4.3 0.6 average 1.0 4.3 4.2 0.6 Table 7 Active ingredients of the glacier water extract of blue hosta gentiana Blue Hosta Gentiana Flower (Glacier Water Extract) Total flavonoids mg / mL Polysaccharide mg / mL Total iridoid glycosides mg / mL Total polyphenols mg / mL Batch 1 1.0 3.9 4.6 0.6 Batch 2 0.9 3.9 4.5 0.5 Batch 3 0.9 4.0 4.6 0.6 average 0.9 3.9 4.6 0.6 It is explained that the blue hosta gentian flower obtained by the blue hosta gentian breeding technology of the present invention is rich in active ingredients and has stable quality. Rich active ingredients can be obtained regardless of pure water extraction or glacial water extraction, and can be developed and applied in the fields of pharmaceuticals, cosmetics, health care products, etc.

[0062] Example 5 Efficacy test of the extract of Hosta gentiana 5.1 Antioxidant efficacy of Gentiana scabra flower extract 1,1-Diphenyl-2-trinitrophenylhydrazine (DPPH) is a stable nitrogen-centered organic free radical. The DPPH method was proposed in 1958 and is widely used to quantitatively determine the anti-aging ability of biological samples, classified substances and foods. This method is based on the characteristics that the DPPH free radical has a single electron, has a strong absorption at 517nm, and its alcohol solution is purple. When a free radical scavenger is present, its absorption gradually disappears due to pairing with its single electron, and its fading degree is quantitatively related to the number of electrons it accepts. Therefore, a spectrophotometer can be used for rapid quantitative analysis to detect the free radical scavenging situation, thereby evaluating the anti-aging ability of the sample.

[0063] Take 0.1 mL of the extract sample solution, add 0.1 mL of DPPH ethanol solution, mix well, react in the dark at room temperature for 30 min, measure the OD value at 525 nm, and calculate the clearance rate according to the following formula: Clearance rate I (%) = [1-(T-T0) / (C-C0)] × 100%, where: T0: absorbance of 0.1 mL of sample solution plus 0.1 mL of 95% ethanol; T: absorbance of 0.1 mL of sample solution plus 0.1 mL of DPPH solution; C0: absorbance of 0.1 mL of water plus 0.1 mL of 95% ethanol; C: absorbance of 0.1 mL of water plus 0.1 mL of DPPH solution. The test results are shown in Table 8.

[0064] Table 8 DPPH scavenging rate of blue hosta gentian flower extract Blue Hosta Gentiana Flower Extract Test concentration (mg / mL) DPPH removal rate of pure water extract (%) DPPH scavenging rate of glacier water extract (%) Batch 1 0.5 83.83 39.43 Batch 2 0.5 87.62 29.71 Batch 3 0.5 87.36 41.25 average 0.5 86.27 36.79 From the results in Table 8, it can be seen that the blue hosta gentian flower extract has a certain effect of scavenging free radicals. The gentian flower extract obtained by pure water extraction and the gentian flower extract extracted by glacier water both have antioxidant effects at 0.5 mg / mL, but the antioxidant effect of the pure water extract is stronger than that of the glacier water extract.

[0065] 5.2 Whitening effect of Gentiana scabra flower extract The physiology, development and metabolism of zebrafish are highly similar to those of mammals, and can reliably simulate human physiology and disease processes. Zebrafish have 87% homology with human genes, and their skin structure and function are very similar to those of humans. More and more people at home and abroad are using zebrafish models to evaluate the efficacy of cosmetics and their raw materials.

[0066] (1) Zebrafish are transparent in the early stages of development, and melanin begins to grow from the retinal epithelium at 24 hours of embryonic development. Pigment cells originate from a group of cells differentiated from the dorsal ectoderm - neural crest cells, which then proliferate, migrate, and differentiate into pigment stem cells. Intervention in the process of melanin formation can inhibit the formation of melanin. The experiment evaluated whether the sample had a whitening effect by analyzing the intensity of the melanin signal on the zebrafish head skin.

[0067] The evaluation method is as follows: Zebrafish skin whiteness evaluation experiment: 6 hpf wild-type AB strain zebrafish were randomly selected in a 6-well plate, with 15 fish in each well; the volume ratio of blue hosta gentian extract to the culture medium was 10%, and it was incubated at 28°C in the dark for 45 hours; 10 zebrafish were randomly selected from each experimental group and placed under a dissecting microscope to take pictures, collect data, and analyze the melanin signal intensity (S) of the zebrafish head. The statistical analysis results of this indicator were used to evaluate the whitening effect of the sample. The inhibition rate of the sample on the melanin signal intensity of the zebrafish head is the whitening effect, and its calculation formula is as follows:

[0068] From Table 9 and Figure 4 As shown, the extract of blue hosta gentian has a certain whitening effect.

[0069] Table 9 Effect of Blue Hosta Gentiana flower extract on zebrafish skin whiteness (2) Intervention in the process of melanin formation can inhibit the formation of melanin. Tyrosinase is the key enzyme in the formation of melanin. The melanin progenitor cells located in the basal layer of the epidermis are activated and secrete melanin. Ultraviolet rays and other factors stimulate melanin and activate the activity of tyrosinase. Tyrosine is converted into DOPA under the action of tyrosinase, and DOPA continues to Dopaquinone is generated under the action of tyrosinase, which is the precursor of pheomelanin. At the same time, dopaquinone generates dopachrome under the catalysis of tyrosinase. Then, under the action of dopachrome tautomerase, most of dopachrome generates dihydroxyindole (DHI) and a small part generates dihydroxyindolecarboxylic acid (DHICA). Under the action of DHICA oxidase, the two generate two kinds of true melanin, DHI-melanin and DHICA-melanin. Therefore, the activity of tyrosinase can be used to evaluate whether the sample has whitening effect.

[0070] Zebrafish tyrosinase activity detection experiment: zebrafish were randomly selected in a 6-well plate, with 30 fish in each well; water-soluble samples were given, and a normal control group was set up at the same time, with a capacity of 3 mL per well. Three biological replicates were performed. Incubate in the dark at 28 °C for 45 hours. After collecting the samples, grind the homogenate with a grinder, centrifuge and take the supernatant, and use the BCA protein concentration assay kit to determine the protein concentration of each experimental group. Take 250 μg of total protein from the zebrafish sample, add 1 mM levodopa solution at a volume ratio of 1:1, and mix well. Transfer the liquid to a 96-well plate, 200 μL / well, and use an enzyme reader to measure the absorbance value at a wavelength of 475 nm. The whitening effect of the sample was calculated according to the formula. The inhibition rate of the sample on zebrafish tyrosinase is the whitening effect. The calculation formula is as follows:

[0071] As shown in Table 10, the blue hosta gentiana flower extract has a certain whitening effect.

[0072] Table 10 Effect of Blue Hosta Gentiana Flower Extract on Tyrosinase Activity project Sample volume Tyrosinase activity inhibition rate (%) pure water extraction Tyrosinase activity inhibition rate (%) Glacier water extract Blue Hosta Gentiana Flower Extract 10% 37 48 (3) Zebrafish are transparent in the early stages of development. Melanin begins to grow from the retinal epithelium at 24 hours of embryonic development. Pigment cells originate from a group of cells differentiated from the dorsal ectoderm - neural crest cells, which then proliferate, migrate and differentiate into pigment progenitor cells. Intervention in the process of melanin formation can inhibit the formation of melanin. The zebrafish melanin protein content was tested to evaluate whether the sample had whitening effect. Zebrafish melanin protein content test: zebrafish were randomly selected in a 6-well plate, 30 per well; water-soluble samples were given, and a normal control group was set up at the same time, with a volume of 3 mL per well. Three biological replicates were performed; incubated at 28 °C in the dark for 45 h. The samples were collected in 1.5 mL centrifuge tubes, RIPA lysis buffer was added to each tube, the slurry was ground with a grinder, and the supernatant was removed by centrifugation. 350 μL of 0.2 M NaOH solution was added to each tube and the tubes were incubated in a water bath at 60 °C for 1 h. Melanin protein standard solution was prepared according to a certain concentration using 0.2 M NaOH solution. The standard solution and the sample to be tested were transferred to a 96-well plate, 100 μL / well, and the absorbance value (OD405) was measured at a wavelength of 405 nm using an enzyme reader. The melanin protein content (C) of each sample group was obtained based on the standard curve. The inhibition rate of the sample on zebrafish melanin protein is the whitening effect. The formula was used to calculate and determine whether the sample has whitening effect.

[0073]

[0074] As shown in Table 11, the blue hosta gentiana flower extract has a certain whitening effect.

[0075] Table 11 Effect of Blue Hosta Gentiana Flower Extract on Melanin Content project Sample concentration Melanin inhibition rate of pure water extract (%) Melanin inhibition rate of glacier water extract (%) Blue Hosta Gentiana Flower Extract 10% 48 66 It can be seen that the gentian flower extract obtained by pure water extraction and the gentian flower extract extracted by glacier water both have whitening effects at a sample concentration of 10%, but the melanin signal intensity inhibition rate, tyrosinase activity inhibition rate, and melanin protein inhibition rate of the glacier water extract are better than those of the pure water extract.

[0076] 5.3 Anti-inflammatory and soothing effects of Gentiana scabra flower extract Sodium dodecyl sulfate (SLS) may cause the body to produce irritation after acting on the body. The irritant enters the body of zebrafish, induces inflammatory response, and neutrophils have immune response, migrate to the skin epidermis and aggregate. TNF-α is the earliest and most important inflammatory cytokine in the inflammatory response process, which can activate neutrophils; IL-1 plays a key role in transmitting information, activating and regulating immune cells, and proliferating and differentiating inflammation in the inflammatory response. IL-1 has two different molecular forms, one is called IL-1α and the other is called IL-1β; IL-16 can promote the upregulation of neutrophil function, promote and regulate the secretion of inflammatory mediators such as cytokines, adhesion molecules and nitric oxide; IL-8 is secreted by monocytes and vascular endothelial cells, and its level rises rapidly when inflammation occurs. The relative expression of zebrafish TNF-α, IL-1α, IL-6 and IL-8 genes can be detected to evaluate whether the sample has a soothing effect.

[0077] Zebrafish were randomly selected in a 6-well plate, with 30 fish per well. SLS was administered in water to establish a zebrafish skin inflammation model. Samples were administered in water, and blank and model control groups were set up at the same time, with a volume of 3 mL per well. Three biological replicates were performed. Incubated in the dark at 28 °C for 18 h. Total RNA of zebrafish in each experimental group was extracted, cDNA was synthesized, and q-PCR was used to detect the gene expression of β-actin and the target gene. β-actin was used as an internal reference for gene expression to calculate the relative RNA expression of the target gene. The blank group was the gene expression level before treatment, and the model group was the gene expression level without the addition of gentian flower extract after stimulation.

[0078] As shown in Table 12, compared with the model group, the blue hosta gentian flower extract down-regulated the relative expression levels of TNF-α, IL-1α, IL-6 and IL-8 genes, indicating that it has certain anti-inflammatory and soothing effects.

[0079] Table 12 Effects of Blue Hosta Gentiana extract on the relative expression levels of TNF-α, IL-1α, IL-6 and IL-8 genes Project Name Sample concentration Blank control group Model Group Pure water extraction Glacier water extraction TNF-α 1% 0.110 1 0.258 0.346 IL-1α 1% 0.472 1 0.768 0.554 IL-6 1% 0.493 1 0.768 0.746 IL-8 1% 0.222 1 0.547 0.561 The gentian flower extract obtained by pure water extraction and the gentian flower extract extracted by glacier water both have anti-inflammatory and soothing effects at a sample concentration of 1%. Pure water is better in down-regulating the gene expression of IL-1α and IL-6, while glacier water is better in down-regulating the gene expression of TNF-α and IL-8.

[0080] In summary, the pure water / glacier water extract of blue hosta gentian has good application prospects in whitening, anti-oxidation, anti-inflammatory and soothing.

[0081] The embodiments described above only express several implementation methods of the present invention and are not intended to limit the scope of protection of the present invention. Those skilled in the art may make various equivalent modifications or substitutions without departing from the creative spirit of the present invention. These all belong to the scope of protection of the present invention and should be subject to the claims.

Claims

1. A method for promoting seed germination and / or rooting of cuttings of Gentiana scabra, characterized in that: The following steps are included: 1) Pre-treating the seeds and / or cuttings of the blue hosta gentian; The pre-treatment is to use glacier water and / or deionized water to treat seeds and / or cuttings, and the treatment time is 20 to 72 hours; 2) Planting the treated blue hosta gentian seeds and / or cuttings into a seedling medium and / or a cutting medium, and controlling the temperature and humidity of the growth environment, thereby promoting the germination of the blue hosta gentian seeds and / or the rooting of the cuttings; The seed germination temperature is 20-30°C, the soil humidity is above 30%, and the air humidity is 60%-70%; The rooting temperature of the cuttings is 18-28° C., and the humidity is above 85%.

2. The method for promoting seed germination and / or rooting of Gentiana scabra as claimed in claim 1, characterized in that: The method for promoting seed germination comprises the following steps: Soak the seeds in glacial water and / or deionized water for 20 to 72 hours. Sow the soaked seeds into the container filled with seedling medium. Cover the top with a layer of vermiculite. The glacier water is collected from the Qudennima Spring in the Himalayas of Tibet. The seedling growing medium comprises fine sand or fine sawdust.

3. The method for promoting seed germination and / or rooting of Gentiana scabra as claimed in claim 1, characterized in that: The seeds are subjected to a germination treatment, wherein the germination treatment comprises the following steps: i) Soak the seeds in 0.05-0.2 ml / L of gibberellin in glacial water and / or deionized water for 20-72 hours, remove and rinse with clean water. ii) Mix the seeds and fine sand evenly, put them into a container, place it in a sunny place, cover the container with wet gauze for germination, control the temperature at 22-25℃, and germinate for 5-7 days.

4. The method for promoting seed germination and / or rooting of cuttings of Gentiana scabra as claimed in claim 1, characterized in that: The cutting rooting process comprises the following steps: Prepare a rooting solution with an IBA content of 5 to 150 mg / kg using glacier water and / or deionized water, and immerse the blue hosta gentian cuttings in the prepared rooting solution for 20 to 72 hours; Insert the pretreated blue hosta gentian cuttings into the cutting medium to promote the rooting of the blue hosta gentian cuttings. Wherein, the glacier water is collected from Qudennima Spring in the Himalayas of Tibet, and the cutting medium contains vermiculite.

5. The method for promoting seed germination and / or rooting of cuttings of Gentiana scabra as claimed in claim 4, characterized in that: The pre-treatment of the cuttings for rooting also includes the following steps: i) Seedling strengthening: Collect blue hosta gentian seedlings and transplant them to the greenhouse for seedling strengthening. The temperature is controlled at 18-28℃ and the humidity is controlled at more than 85%. Plant for more than 10 days; ii) Cut the cuttings from the blue hosta gentian seedlings, select strong and disease-free tender branches, cut them into 2-3 cm cuttings at the lower part of the branches, make the cuts smooth, remove the base leaves, and trim the upper leaves of the cuttings appropriately; iii) Sterilize the base of the cuttings with 0.03% KMnO4 for 5 to 10 minutes.

6. An ecological cultivation method of blue hosta gentian, comprising the following steps: 1) Promoting the germination of blue hosta gentian seeds and / or rooting of cuttings by the rooting method according to claims 1 to 5 to obtain rooted blue hosta gentian seedlings; 2) Seedling cultivation, the seedling cultivation process includes the following steps: The gentian seedlings are transplanted into a seedling tray containing a culture medium, The gentian seedlings were sprayed with fertilizer, the fertilizer was "Pendefu", the application concentration was 400-1200 times, and the vermiculite surface was sterilized with 0.03% KMnO4 solution every half a month after the seedlings emerged. The cultivation conditions for the seedling cultivation are room temperature of 18-28°C, humidity of more than 85%, shade, and watering 2-3 times a week. The cultivation matrix is ​​a mass ratio of perlite: garden soil = 0.5-1.5: 1.5-2.5, the soil binder is HPMC, the concentration of the HPMC is 200-400 mg / L, and the pH of the cultivation matrix is ​​5.7-6.79; 3) Large-scale transplanting The gentian seedlings are transplanted to the field after growing for one year, and the transplanting time is in early to mid-April in spring or in late September to early October in autumn. Before transplanting, the seedlings are placed outside the greenhouse for 7 to 10 days of seedling hardening. The planting method is as follows: covering the ground with a film, drilling holes for planting, planting 2 to 3 gentian seedlings in each hole, and the planting spacing is greater than 20 cm. During transplanting, continuous spraying is performed for at least 3 days, and before flowering, in mid-to-late July, fertilizer containing a humic acid aqueous solution is applied.

7. The ecological cultivation method according to claim 6, characterized in that: The mass ratio of the perlite to the garden soil is 1:2, and the concentration of the HPMC is 300 mg / L.

8. A method for preparing a blue hosta gentiana flower extract, characterized in that: The steps include:

1. Collect blooming blue hosta gentiana flowers, dry them at 35-45°C, crush them and sieve them to obtain blue hosta gentiana flower powder; 2. Take the blue hosta gentian flower powder, add the solvent at a solid-liquid ratio of 1:10-100, perform ultrasonic extraction at 20-50°C with an ultrasonic power of 400-700W for more than 30 minutes, and then centrifuge and filter at 10000rpm to obtain the blue hosta gentian flower extract; Further, the solvent is one or more of glacier water, deionized water, ethanol, butanediol, and further, the solvent is deionized water or glacier water. The material-liquid ratio is preferably 1:20, 1:50, and the ultrasonic power is 600W.

9. A skin external preparation with anti-aging, whitening, anti-inflammatory, soothing, sunscreen or antioxidant functions, characterized in that: The active ingredient is blue hosta gentian flower extract.

10. A skin topical preparation having the functions of scavenging free radicals, inhibiting tyrosinase activity, inhibiting melanin production, preventing oxidative damage, and inhibiting TNF-α, IL-1α, IL-6 and / or IL-8 gene expression, characterized in that: The active ingredient is blue hosta gentian flower extract.

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