Dendrobium nobile fertilizer as well as fermentation method and application thereof

Abstract

The invention discloses a dendrobium nobile fertilizer and a fermentation method and application thereof.The dendrobium nobile fertilizer comprises a vinasse matrix and gneiss, the mass ratio of the vinasse matrix to the gneiss is (15-20): 1, aired and smashed vinasse and rape seed cakes are mixed according to the mass ratio, then em bacterial powder is added, fermentation is conducted for 15-30 days, turning is conducted 2-4 times during fermentation, and the fermentation time is 3-5 days; turning over when the fermentation temperature is higher than 65 DEG C, controlling the temperature to be below 65 DEG C, and stopping fermentation until the fermentation peculiar smell is weakened and the temperature is reduced to room temperature, so as to prepare a vinasse substrate; the invention further discloses application of the dendrobium nobile fertilizer in dendrobium nobile cultivation and planting. The prepared dendrobium nobile fertilizer can be used as a good substitute material for non-renewable resources such as pine barks, high-value development and utilization of vinasse waste resources are achieved, the epigenetic characteristics of provenance can be well reserved when dendrobium nobile is cultivated through the vinasse substrate, the cultivated dendrobium nobile grows normally in the growth cycle, and the yield of dendrobium nobile is increased. The quality of the harvested dendrobe meets the national standard, and the method has a good application effect.

Description

Technical Field

[0001] The present invention relates to the technical field of Dendrobium seedling cultivation, and particularly relates to a Dendrobium fertilizer, a fermentation method thereof, and an application thereof. Background Art

[0002] Huoshan Dendrobium is a herbaceous plant under the genus Dendrobium in Orchidaceae, and can also be called "Mihu". Its original place is the Dabie Mountain area in Anhui Province. It is a rare local medicinal material in Anhui Province. The pharmacological active ingredients include polysaccharides, alkaloids, amino acids, and flavonoids, and it has various pharmacological effects such as immune regulation, anti-tumor, and antioxidant. Currently, it has become an important research object in aspects such as traditional Chinese medicine and health food. Because it mainly grows in the stone crevices of cliffs, with a too long growth cycle and a high dependence on the growth environment, and coupled with over-harvesting by humans, it has been on the verge of extinction several times.

[0003] In recent years, due to the off-site cultivation and facility breeding of traditional Chinese medicines, the phenomenon of reduced quality has become increasingly prominent, which has also led to the general preference for cultivation methods that return to the original place and the original ecological environment. At present, a series of research results have been obtained in the field of Dendrobium in China. For example, Wu Tingting et al. studied the influence of exogenous organic carbon on the accumulation of internal substances in Dendrobium officinale, and found α-ketoglutaric acid, which can promote its growth and development and improve the quality. In terms of Huoshan Dendrobium, Yi Shanyong et al. studied the comprehensive benefits of three planting modes, namely facility cultivation, under-forest cultivation, and simulated environment cultivation in the Dabie Mountain area, and found that the simulated environment cultivation has the highest medicinal components and economic benefits and is most suitable for large-scale planting; Wang Xiusong et al. specified the wild-simulated cultivation technology of Huoshan Dendrobium from aspects such as forest land selection, infrastructure construction, seedling selection, cultivation, and management, providing a reference for high-quality cultivation; Wang Zhaocheng et al. determined the growth amount, polysaccharides, alkaloids, amino acids, and moisture content of Huoshan Dendrobium under three cultivation modes, namely greenhouse cultivation, forest gap cultivation, and living tree epiphytic cultivation, and compared the differences in their growth and internal substances to clarify the suitable cultivation mode, and concluded that the Huoshan Dendrobium under living tree epiphytic cultivation accumulates more active ingredients and has better quality, which is a planting method suitable for popularization in the Dabie Mountain area. As an endangered traditional Chinese medicine resource, Huoshan Dendrobium has mainly been replaced by cultivation instead of wild picking for many years. Now, people can use different substrates and cultivation modes to preserve this species of Huoshan Dendrobium. Currently, the commonly used planting substrates are mixtures of wood resources such as pine bark and stones. However, the current wood matrix resources such as pine bark are limited, the wood resources are exhausted, and the cultivation cost increases. It is necessary to select more suitable planting substrates to reduce the cultivation cost while maintaining the original quality.

[0004] Distillers' grains, also known as spent grains, are the solid mixtures remaining after distilling alcohol from the raw materials of corn, sorghum, wheat and other grains, using distiller's yeast, saccharifying enzyme, etc. as fermentation agents through steaming, saccharification fermentation. They are the main by-products of the alcohol-making industry. Rich in organic matter, distillers' grains are a high-quality biomass resource, and their resource utilization is of great significance for environmental protection and the green, low-carbon and circular production of the brewing industry. They are commonly used in livestock breeding, production fermentation and the cultivation of fruits, vegetables and melons. At present, domestic scholars have carried out a number of studies on the reuse of distillers' grains. For example, Liu Yunfei et al. summarized the effects of current liquor spent grains as feed on the production performance of cattle, pigs and sheep; Liu Liancheng used the mixture of distillers' grains and cottonseed hulls as the carbon source matrix of Hericium erinaceus, and studied the effects of different formulations on the mycelial growth status and the yield of fresh Hericium erinaceus, providing a scientific basis for the selection of cultivation raw materials for Hericium erinaceus and the comprehensive utilization of distillers' grains. Zhang Jinghui et al. combined distillers' grains and oil-tea camellia shells for edible mushroom cultivation to explore the best plan for the compatibility cultivation of distillers' grains and oil-tea camellia shells with edible mushrooms; Wan Hefeng et al. carried out composting treatment on blueberry wine distillers' grains and cow dung in a certain proportion, and through in-situ monitoring, sampling analysis and comparison with two commercial substrates A and B for blueberry cultivation, evaluated the suitability of blueberry fermented distillers' grains as a blueberry cultivation substrate after composting treatment. Therefore, distillers' grains are a relatively ideal alternative substrate, and at present, some enterprises have explored using resources such as distillers' grains to replace bark as a planting substrate.

[0005] Different cultivation substrates have a direct impact on the main component content and medicinal efficacy of plants. Hou Yuhao et al. explored the effects of different origins, cultivation modes and cultivation substrates on the main component content and in vitro anti-tumor activity of Phellinus igniarius, and concluded that the main component content and activity of Phellinus igniarius are affected by varieties, cultivation modes and substitute material cultivation substrates, and the material basis for the in vitro anti-tumor activity of Phellinus igniarius is mainly flavonoids and total phenols, and high polysaccharide content will affect the in vitro anti-tumor activity; Zhang Shuyu et al. explored the effects of waste compost as a cultivation substrate for Dianthus chinensis, Viola cornuta and Bellis perennis in order to make resource utilization of garden waste. Similarly, different cultivation substrates have a certain impact on the growth status and even endophytic fungi of Dendrobium huoshanense. Then, what is the impact of using distillers' grains substrate to replace pine bark substrate on Dendrobium, whether the quality of the medicinal materials obtained by cultivating with distillers' grains as the substrate is the same, and whether there are compositional differences between the two have not been demonstrated. Summary of the Invention

[0006] The purpose of the present invention is to provide a Dendrobium fertilizer, its fermentation method and application to solve the problems raised in the above background technology.

[0007] To achieve the above purpose, the present invention is realized through the following technical means:

[0008] A Dendrobium fertilizer includes distillers' grains substrate and gneiss, and the mass ratio of the distillers' grains substrate to gneiss is 15 - 20:1.

[0009] Among them, the distillers' grains matrix includes distillers' grains and rapeseed cakes, and the mass ratio of distillers' grains to rapeseed cakes is 13 - 30:1, and the mass fraction of 0.1 - 0.5% of EM bacteria powder is 1% - 2%.

[0010] Furthermore, the distillers' grains need to be sun-dried for 2 - 3 days to allow part of the alcohol to volatilize, reducing the damage of alcohol to the roots of Dendrobium, and the sun-dried distillers' grains and rapeseed cakes are crushed into small particles.

[0011] Furthermore, the distillers' grains selected are the distillers' grains discarded after steaming and brewing with sorghum, glutinous rice, rice, wheat, and corn.

[0012] A fermentation method for a Dendrobium fertilizer includes the following steps:

[0013] 1), Mix the sun-dried and crushed distillers' grains with rapeseed cakes according to the mass ratio, then add EM bacteria powder and ferment for 15 - 30 days, during which it is turned over 2 - 4 times. When the fermentation temperature is higher than 65°C, it is turned over to keep the temperature below 65°C until the fermentation odor fades and the temperature drops to room temperature, then stop fermentation to make the distillers' grains matrix;

[0014] 2), Wash and remove the sediment from the gneiss with a particle size of 2 - 3 cm, take the fermented distillers' grains matrix, and stir evenly according to the mass ratio of matrix:gneiss to obtain the mixed distillers' grains matrix;

[0015] 3), Place 2.5 kg of gneiss with a particle size of 6 cm at the bottom of the seedling tray, spread the mixed distillers' grains matrix on top of the gneiss, making it 1 cm higher than the top of the seedling tray. Water it thoroughly before planting, and then enter the planting process.

[0016] Furthermore, keep the water content of the matrix at 50% - 60%, that is, it forms a ball when held in the hand and disperses when released. Stack the mixed matrix in a well-ventilated place away from direct sunlight. The state of fermentation completion is: when the color of the material becomes darker, there is no odor, and the texture is soft.

[0017] An application of a Dendrobium fertilizer in the cultivation of Dendrobium huoshanense.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] The Dendrobium fertilizer prepared by the present invention can be used as a good alternative material for non-renewable resources such as pine bark. At the same time, the waste resources of distillers' grains are highly valorized and developed. Using the distillers' grains matrix of the present invention to cultivate Dendrobium can well retain the epigenetic characteristics of the germplasm source. The cultivated Dendrobium grows normally during the growth cycle, and the quality of the harvested Dendrobium meets the national standards. The distillers' grains cultivation matrix of the present invention has good application effects. Description of the Drawings

[0020] Figure 1 Schematic diagram of the application of the distiller's grains substrate of the present invention in the cultivation of Dendrobium huoshanense

[0021] Figure 2 Schematic diagram for comparison of planting with the 1# source distiller's grains substrate (left) and planting with the 1# source pine bark substrate (right) in the present invention

[0022] Figure 3 Schematic diagram for comparison of planting with the 2# source pine bark substrate (left) and planting with the 2# source distiller's grains substrate (right) in the present invention

[0023] Figure 4 Schematic diagram of the influence of the cultivation substrate on the crude polysaccharide content of Dendrobium huoshanense in the present invention Detailed implementation manners

[0024] The embodiments of the technical solutions of the present application will be described in detail below with reference to the accompanying drawings. The following embodiments and the accompanying drawings are only used to illustrate the technical solutions of the present application more clearly, so they are only examples and cannot be used to limit the protection scope of the present application. Only the parts related to the technical solutions of the present application are schematically shown in the accompanying drawings, and they do not represent the actual structure of the product.

[0025] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the technical field to which this application belongs; the terms used herein are only for the purpose of describing specific embodiments and are not intended to limit this application; the terms "including" and "having" and any variations thereof in the description of the specification and claims of this application and the above accompanying drawings are intended to cover non-exclusive inclusion.

[0026] In the description of the embodiments of the present application, technical terms such as "first" and "second" are only used to distinguish different objects and cannot be understood as indicating or implying relative importance or implicitly indicating the quantity, specific order or primary-secondary relationship of the indicated technical features. In the description of the embodiments of the present application, the term "plurality" means two or more (including two). Similarly, "multiple groups" means two or more groups (including two groups), and "multiple pieces" means two or more pieces (including two pieces).

[0027] Referring to "embodiments" herein means that the specific features, structures or characteristics described in connection with the embodiments can be included in at least one embodiment of this application. The phrase appears in various places in the specification and does not necessarily refer to the same embodiment, nor is it an independent or alternative embodiment mutually exclusive with other embodiments. Those skilled in the art explicitly and implicitly understand that the embodiments described herein can be combined with other embodiments.

[0028] In the description of the embodiments of the present application, the term "and / or" is merely an association relationship describing associated objects, indicating that there can be three relationships. For example, A and / or B can represent: A exists alone, A and B exist simultaneously, and B exists alone. Additionally, the character " / " in this text generally indicates an "or" relationship between the associated objects before and after.

[0029] In this embodiment, a Dendrobium fertilizer includes distiller's grains substrate and gneiss, and the mass ratio of the distiller's grains substrate to gneiss is 15 - 20:1.

[0030] Among them, the distiller's grains substrate includes distiller's grains and rapeseed cake, the mass ratio of distiller's grains to rapeseed cake is 13 - 30:1, and the mass fraction of 0.1 - 0.5% EM bacterial powder is 1% - 2%.

[0031] Further, the distiller's grains need to be sun-dried for 2 - 3 days to allow part of the alcohol to volatilize, reducing the harm of alcohol to the roots of Dendrobium, and then the sun-dried distiller's grains and rapeseed cake are crushed into small granular shapes.

[0032] Further, the distiller's grains selected are the distiller's grains discarded after steaming and brewing with sorghum, glutinous rice, rice, wheat, and corn.

[0033] A fermentation method for a Dendrobium fertilizer includes the following steps:

[0034] 1), Mix the sun-dried and crushed distiller's grains with rapeseed cake according to the mass ratio, then add EM bacterial powder and ferment for 15 - 30 days, turning 2 - 4 times during the period. When the fermentation temperature is higher than 65°C, turn to keep the temperature below 65°C until the fermentation odor fades and the temperature drops to room temperature, then stop fermentation to make the substrate;

[0035] 2), Wash and remove the sediment from the gneiss with a particle size of 2 - 3 cm, take the fermented distiller's grains substrate, and stir evenly according to the mass ratio of substrate:gneiss to obtain the mixed distiller's grains substrate;

[0036] 3), Place 2.5 kg of gneiss with a particle size of 6 cm at the bottom of the seedling tray, spread the mixed distiller's grains substrate on top of the gneiss, making it 1 cm higher than the top of the seedling tray. Water thoroughly before planting, and then enter the planting process.

[0037] Further, keep the water content of the substrate at 50% - 60%, that is, it forms a ball when held in the hand and disperses when released. Stack the mixed substrate in a well-ventilated place away from direct sunlight. The state of fermentation completion is: when the color of the material becomes darker, there is no odor, and the texture is soft.

[0038] An application of a Dendrobium fertilizer in the cultivation of Huoshan Dendrobium.

[0039] The Dendrobium officinale from Huoshan from different sources were planted in homemade distiller's grains substrate and traditional pine bark substrate respectively. After three years of cultivation, the growth of the two plants was compared. It was found that there was no significant difference in the appearance of Dendrobium officinale from Huoshan from different sources when they were planted in homemade distiller's grains substrate and traditional pine bark substrate. Figure 2 , Figure 3 The typical "grasshopper leg" phenotype of Zhonghuoshan Dendrobium is obvious.

[0040] In order to be a good substitute material for non-renewable resources such as pine bark, the distiller's grains in the present invention must effectively ensure the components required for the growth of Dendrobium huoshanense, so as to promote the growth of the rhizomes of Dendrobium huoshanense and ensure the quality of Dendrobium huoshanense.

[0041] The present invention discloses an example of analyzing the components of the vinasse matrix.

[0042] Sample source: Huoshan County, Anhui Province, the authentic production area of ​​Huoshan Dendrobium, and Anhui Dabieshan Huohu Technology Co., Ltd. Collection time: November 2023. Samples were collected from the greenhouse cultivation distiller's grains matrix soil of the Huoshan Dendrobium base, and three soils of different depths were taken. The six groups of samples were sealed and preserved, and all matrix samples were transferred by dry ice and stored in an ultra-low temperature refrigerator at -80°C for subsequent testing.

[0043] 1. Determination of water content of vinasse matrix by drying method

[0044] Weigh 5g of the mixed soil sample and put it into a sterilized evaporating dish, then dry it in an oven for 12 hours (temperature range: 104℃~106℃), take it out and put it into a desiccator, cool it to room temperature and weigh it, then dry it for 1 hour, and finally dry it in a desiccator for 30 minutes. Repeat the above process until constant weight is reached.

[0045] 2. Determination of pH of lees matrix by conductivity measurement

[0046] Weigh 10 g of the mixed soil sample, pass it through a 10-mesh sieve, place the sieved sample in a 50 mL beaker, and add deionized water (without CO 2 ), stir thoroughly with a glass rod and let stand for 30 minutes, and finally measure the pH value with a pH conductivity meter.

[0047] 3. Determination of organic matter in vinasse matrix by Qiulin method

[0048] The soil organic matter was determined with reference to NY / T 525-2021. Take 0.4 g of the dried and pulverized soil sample, filter it through a 100-mesh sieve, pour it into a test tube, add 5 mL of 0.8 mol / L potassium dichromate standard solution and 5 mL of sulfuric acid, mix well, place the test tube on a test tube rack and heat it in a water bath for 30 min. After taking it out, let it cool naturally to room temperature. Pour the cooled content of the test tube into a 250 mL Erlenmeyer flask, add 3-5 drops of o-phenanthroline indicator, and titrate with ferrous sulfate solution. During the titration, the solution changes from orange to green, turns dark green when approaching the end point, and finally turns brick red, indicating that the titration reaches the end point.

[0049] IV. Determination of total nitrogen in the distiller's grains matrix by Kjeldahl digestion method

[0050] The total nitrogen was determined with reference to NY / T 53-1987. Weigh 0.1 g of the mixed soil sample, pass it through a 100-mesh sieve, put it into a digestion tube, then add 1 mL of potassium permanganate solution and 2 mL of sulfuric acid solution (the ratio of sulfuric acid to water is 1:1), mix well, and digest it with an electric heating digestion furnace. After completion, transfer it to the distillation tube of an automatic Kjeldahl nitrogen analyzer, add 25 mL of 400 g / L NaOH solution, start to introduce steam, and use H 2 BO 3 solution (20 g / L, 10 mL) as the receiving solution at the bottom of the condenser, and titrate with sulfuric acid standard solution.

[0051] V. Determination of available phosphorus in the distiller's grains matrix soil by ultraviolet spectrophotometry

[0052] The available phosphorus was determined with reference to NY / T 1121.7-2014. Take 2.5 g of the mixed soil sample, pass it through a 10-mesh sieve, pour the sieved sample into a 250 mL conical flask, add 50 mL of NaHCO 3 solution (pH value is 8.5, concentration is 0.5 mol / L), use a shaker to shake for 30 min, set the parameter to 180 r / min, filter after shaking, take 10 mL of the filtrate, add 20 ml of water and 1 drop of dinitrophenol indicator, adjust the pH to colorless, then add 0.75 mL of ascorbic acid and 5 mL of molybdate, make up the volume to 50 mL, let it stand at room temperature for half an hour, and then colorimetric with an ultraviolet spectrophotometer.

[0053] VI. Determination of available potassium in the distiller's grains matrix by flame spectrophotometry

[0054] The available potassium was determined with reference to NYIT 889-2004. Weigh 2.5 g of the mixed soil sample, sieve it through a 10-mesh sieve, then air-dry it, place it in a plastic bottle, add ammonium acetate extraction solution, use a shaker to shake it, set the parameters as constant temperature at 25 °C and 180 r / min, shake for 30 min, filter after shaking, take 5 mL of the filtrate and make up the volume to 50 mL in a volumetric flask, and directly determine it using a flame spectrophotometer.

[0055] The determination results of the water content, pH value and components of the distiller's grains substrate are shown in Table 1.

[0056] Table 1 Determination results of the water content, pH value and components of the distiller's grains substrate

[0057]

[0058] The results show that the distiller's grains substrate is rich in organic matter, nitrogen, phosphorus and potassium, and the water content and pH are appropriate, which can meet the cultivation requirements of Dendrobium officinale.

[0059] VII. Growth situation of Dendrobium huoshanense planted with the distiller's grains substrate of the present invention

[0060] Randomly select 30 plants of Dendrobium huoshanense, use a vernier caliper to measure and record indicators such as stem length (cm), stem diameter (cm), number of nodes (cm), etc. The specific measurement standards are as follows:

[0061] Stem length: Use a meter ruler to measure the total length from the base of the stem to the top of the stem (removing the top leaves), take the highest stem in the fixed-point measurement cluster as the standard, from the base of the stem to the tip; Stem diameter: Use a vernier caliper to measure the thickest part of the stem of Dendrobium huoshanense.

[0062] Collect the plants of Dendrobium huoshanense from different provenances cultivated in pine bark substrate and distiller's grains substrate for 1, 2, and 3 years respectively, remove the roots and leaves, and measure the stem length of the plants. The results show (Table 2) that for Dendrobium huoshanense of the same variety and the same cultivation substrate, the stem length of the 1-year-old plants > the stem length of the 2-year-old plants > the stem length of the 3-year-old plants, which conforms to the growth trend of Dendrobium huoshanense. Comparing different cultivation substrates for the same variety, it is found that the Dendrobium huoshanense planted with the distiller's grains substrate has a shorter stem, and the phenotype is closer to the short and small characteristics of wild Dendrobium huoshanense plants.

[0063] Table 2 Comparison of the stem length of Dendrobium huoshanense in different cultivation substrates (cm, )

[0064]

[0065]

[0066] Plants of Dendrobium huoshanense from different provenances cultivated in pine bark substrate and distiller's grains substrate for 1, 2, and 3 years were collected respectively, with roots and leaves removed, and the stem diameters of the plants were measured. The results showed (Table 3) that there were slight differences in the stem diameters of different varieties of Dendrobium huoshanense. The stem diameters of two-year-old and three-year-old plants were greater than those of one-year-old plants. For Dendrobium huoshanense of the same provenance planted in distiller's grains substrate and pine bark substrate respectively, there were no significant differences in their stem diameters.

[0067] Table 3 Comparison of stem diameters of Dendrobium huoshanense in different cultivation substrates (cm, )

[0068]

[0069] Plants of Dendrobium huoshanense from different provenances cultivated in pine bark substrate and distiller's grains substrate for 1, 2, and 3 years were collected respectively, with roots and leaves removed, and the number of stem nodes of the plants was recorded. The results showed (Table 4) that for Dendrobium huoshanense cultivated in pine bark and distiller's grains substrates of various provenances, the number of stem nodes of one-year-old plants > the number of stem nodes of two-year-old plants > the number of stem nodes of three-year-old plants. There were no significant differences in the number of stem nodes of Dendrobium huoshanense cultivated in pine bark and distiller's grains substrates.

[0070] Table 4 Comparison of the number of stem nodes of Dendrobium huoshanense in different cultivation substrates

[0071]

[0072] The above results showed that Dendrobium huoshanense cultivated in distiller's grains substrate grew normally during the 3-year growth cycle and met the requirements for planting and harvesting. In addition, compared with Dendrobium huoshanense cultivated in pine bark substrate, there were no differences in the stem diameters of Dendrobium huoshanense cultivated in distiller's grains substrate, and the stem lengths were shorter, and the morphology was closer to that of the wild and authentic Dendrobium huoshanense "grasshopper leg", which better met the "excellent shape" requirements of D. huoshanense (commonly known as "Mi Hu").

[0073] VIII. Quality evaluation of Dendrobium huoshanense planted with the distiller's grains substrate of the present invention

[0074] 1.1 Determination of crude polysaccharide content: Phenol-sulfuric acid method

[0075] It was determined by ultraviolet-visible spectrophotometry (General Rule 0401) with reference to the Chinese Pharmacopoeia 2020 Edition.

[0076] 1.1.1 Preparation of reference substance solution: An appropriate amount of D-anhydroglucose reference substance was accurately weighed and dissolved in water to make a solution containing 100 μg per 1 ml, and that was obtained.

[0077] 1.1.2 Preparation of Standard Curve Precisely measure 0.2 mL, 0.4 mL, 0.6 mL, 0.8 mL, and 1.0 mL of the reference substance solution respectively, place them in 10-mL stoppered test tubes, add water to each tube to make up to 1.0 mL, precisely add 1 mL of 5% phenol solution (prepared immediately before use), shake well, then precisely add 5 mL of sulfuric acid, shake well, place in a boiling water bath and heat for 20 minutes, take out, cool in an ice bath for 5 minutes. Using the corresponding reagent as the blank, according to the ultraviolet-visible spectrophotometry (General Principles 0401), measure the absorbance at a wavelength of 488 nm. Taking the absorbance as the ordinate and the concentration as the abscissa, plot the standard curve.

[0078] 1.1.3 Determination Method Take about 0.4 g of the powder of this product (the fresh product is dried and pulverized, passing through No. 3 sieve), precisely weigh it, add 200 mL of water, heat under reflux for 2 hours, cool, transfer to a 250-mL volumetric flask, wash the container with a small amount of water several times, combine the washings into the same volumetric flask, add water to the scale, shake well, and filter. Precisely measure 2 mL of the consecutive filtrate, place it in a 15-mL centrifuge tube, precisely add 10 mL of absolute ethanol, shake well, refrigerate for 1 hour, take out, centrifuge (at a speed of 4000 revolutions per minute) for 20 minutes, discard the supernatant, wash the precipitate with 80% ethanol twice, 8 mL each time, centrifuge, discard the supernatant, dissolve the precipitate by heating with hot water, transfer to a 25-mL volumetric flask, let it cool to room temperature, add water to the scale, and shake well. Precisely measure 1 mL of the test solution, place it in a 10-mL stoppered test tube, according to the method under the preparation of the standard curve, starting from "precisely add 1 mL of 5% phenol solution", measure the absorbance according to the law, read the amount of D-anhydroglucose in the test solution from the standard curve, calculate, and you will get the result.

[0079] 1.2 Determination of Extract Content

[0080] For Dendrobium huoshanense, determine by the hot extraction method under the determination method of alcohol-soluble extracts (General Principles 2201). Use ethanol as the solvent. Take about 2 - 4 g of the test sample, precisely weigh it, place it in a 100 - 250-mL conical flask, precisely add 50 - 100 mL of ethanol, tightly stopper, weigh, let stand for 1 hour, connect a reflux condenser, heat to boiling, and keep slightly boiling for 1 hour. After cooling, remove the conical flask, tightly stopper, weigh again, make up the lost weight with ethanol, shake well, filter through a dry filter. Precisely measure 25 mL of the filtrate, place it in an evaporating dish that has been dried to a constant weight, evaporate to dryness on a water bath, dry at 105 °C for 3 hours, cool in a desiccator for 30 minutes, and quickly and precisely weigh. Unless otherwise specified, calculate the content (%) of water-soluble extracts in the test sample based on the dried product.

[0081] 2 Test Results 2.1 Influence of Cultivation Substrate on the Content of Crude Polysaccharides in Dendrobium huoshanense Respectively take 10 clusters of Dendrobium huoshanense plants cultivated in pine bark substrate and distiller's grains substrate, remove the roots and leaves, dry them, precisely weigh the dry products of Dendrobium huoshanense in each group, conduct sample treatment according to the preparation method of test samples, establish a glucose standard curve, and obtain the curve equation as y = 10.704x - 0.00587, R2 = 0.9998. Substitute the absorbance into the standard curve to calculate the crude polysaccharide content of Dendrobium huoshanense in each group.

[0084] Table 6 Comparison of the crude polysaccharide content of Dendrobium huoshanense in different cultivation substrates (calculated based on dry products, %)

[0085]

[0086] The results show that the total polysaccharide content of Dendrobium huoshanense cultivated in distiller's grains substrate is slightly lower than that of the same-source Dendrobium huoshanense cultivated in pine bark substrate, but there is no significant difference between the two. Moreover, the requirements under Dendrobium huoshanense in the Chinese Pharmacopoeia state that for Dendrobium huoshanense calculated based on the dry product, the polysaccharide content calculated as anhydrous glucose shall not be less than 17%. The total polysaccharide content in the dry products of Dendrobium huoshanense cultivated in distiller's grains substrate all meets the requirements of the Chinese Pharmacopoeia.

[0087] 2.2 Influence of cultivation substrate on the extract content of Dendrobium huoshanense

[0088] Respectively take 10 clusters of Dendrobium huoshanense plants cultivated in pine bark substrate and distiller's grains substrate, remove the roots and leaves, dry them, precisely weigh the dry products of Dendrobium huoshanense in each group, conduct sample treatment according to the preparation method of test samples, determine the content of alcohol-soluble extracts, and calculate the extract yield.

[0089] Table 7 Extract yield of alcohol-soluble extracts of Dendrobium huoshanense in different cultivation substrates (calculated based on dry products, %)

[0090]

[0091] The requirements under Dendrobium huoshanense in the Chinese Pharmacopoeia state that according to the hot extraction method under the determination method of alcohol-soluble extracts (General Rule 2201), using ethanol as the solvent, the dry product shall not be less than 8.0%. The above results show that the extract content in the dry products of Dendrobium huoshanense cultivated in distiller's grains substrate meets the requirements under Dendrobium huoshanense in the Chinese Pharmacopoeia. Moreover, the content of alcohol-soluble extracts of Dendrobium huoshanense cultivated in distiller's grains substrate is slightly higher than that of the same-source Dendrobium huoshanense cultivated in pine bark substrate, but there is no significant difference between the two. The quality of Dendrobium huoshanense cultivated in distiller's grains substrate meets the requirements of the Chinese Pharmacopoeia, and this distiller's grains substrate can be used as a substitute for the non-renewable pine bark resource for Dendrobium cultivation.

[0092] In summary, when using the distiller's grains substrate of the present invention to cultivate Dendrobium officinale, it can well retain the epigenetic characteristics of the germplasm source. The cultivated Dendrobium officinale grows normally during the growth cycle, and the quality of the harvested Dendrobium officinale meets the national standards. Therefore, the distiller's grains cultivation substrate in the present invention has good application effects, can be used as a good alternative material for non-renewable resources such as pine bark, and at the same time realizes the high-value development and utilization of distiller's grains waste resources.

[0093] The specific embodiments disclosed in the present invention fall within the protection scope of the claims of the present invention, which is the specific lower-level implementation scope of the feature part of the present invention. The protection content of the specific embodiments is only an explanation of the protection scope of the claims of the present invention. The protection scope of the present invention is not limited to the protection content of the specific embodiments, and the protection content of the specific embodiments should not be construed as a limitation on the protection scope of the claims of the present invention.

[0094] Only some exemplary embodiments of the present invention have been described by way of illustration above. Without doubt, for those of ordinary skill in the art, the described embodiments can be modified in various different ways without departing from the scope of the present invention. Therefore, the above description is illustrative in nature and should not be construed as a limitation on the protection scope of the claims of the present invention.

[0095] Unless otherwise defined, all academic and scientific terms used herein have the same meaning as understood by those of ordinary skill in the technical field to which the present invention pertains.

[0096] In case of conflict, the definitions in this specification shall prevail.

[0097] Unless otherwise specified, all percentages, parts, ratios, etc. are by weight.

[0098] When a numerical value or range of values, a preferred range, or a series of lower preferred values and upper preferred values are given, it should be understood that any range formed by any pair of numerical values consisting of any smaller range limit value or preferred value and any larger range limit value or preferred value is specifically disclosed, regardless of whether the ranges are separately disclosed. Unless otherwise specified, where a numerical range is described in this specification, the range is intended to include the range end values and all integers and fractions within the range.

[0099] When the terms "about" or "around" are used to describe the end values of a numerical value or range, the disclosed content should include the specific numerical value or the end value involved.

[0100] The use of the articles "a" and "one / kind" to describe the elements of the present invention is only for convenience and to give a general description of the present invention. Unless otherwise clearly stated, this description should be understood to include one / kind or at least one / kind.

Claims

1. A dendrobium fertilizer, characterized in that: The invention comprises a wine lees matrix and gneiss, wherein the mass ratio of the wine lees matrix to the gneiss is 15-20:

1.

2. A Dendrobium fertilizer according to claim 1, characterized in that: The vinasse matrix comprises vinasse and rapeseed cake, the mass ratio of vinasse to rapeseed cake is 13-30:1, and the mass fraction of 0.1-0.5% EM bacterial powder is 1%-2%.

3. The dendrobium fertilizer according to claim 1, characterized in that: The lees need to be placed in the sun for 2-3 days to allow some of the alcohol to evaporate and reduce the damage of the alcohol to the root system of Dendrobium, and the lees and rapeseed cake after drying are crushed into small particles.

4. The dendrobium fertilizer according to claim 1, characterized in that: The selected distiller's grains are the distiller's grains discarded after sorghum, glutinous rice, rice, wheat and corn are cooked and made into wine.

5. A fermentation method for dendrobium fertilizer, characterized in that: The following steps are included: 1) Mix the dried and crushed distiller's grains with rapeseed cake according to the mass ratio, then add EM powder, ferment for 15-30 days, turn over 2-4 times during the fermentation, turn over when the fermentation temperature is higher than 65°C, control the temperature below 65°C, until the fermentation odor becomes lighter and the temperature drops to room temperature, stop fermentation, and prepare distiller's grains matrix; 2) Wash the gneiss with a particle size of 2-3 cm to remove the sand, take the fermented distiller's grains matrix, and stir evenly according to the mass ratio of matrix: gneiss to prepare a mixed distiller's grains matrix; 3) Place 2.5 kg of gneiss with a particle size of 6 cm at the bottom of the seedling tray, spread the mixed wine lees matrix on the gneiss to make it 1 cm higher than the top of the seedling tray, water it thoroughly before planting, and then enter the planting stage.

6. Use of the dendrobium fertilizer as described in claims 1-4 in dendrobium cultivation.

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