CN109819868B - Dendrobium officinale culture medium and preparation method thereof - Google Patents
Dendrobium officinale culture medium and preparation method thereof Download PDFInfo
- Publication number
- CN109819868B CN109819868B CN201910280356.8A CN201910280356A CN109819868B CN 109819868 B CN109819868 B CN 109819868B CN 201910280356 A CN201910280356 A CN 201910280356A CN 109819868 B CN109819868 B CN 109819868B
- Authority
- CN
- China
- Prior art keywords
- dendrobium officinale
- culture medium
- parts
- cfu
- preparation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Abstract
The invention discloses a dendrobium officinale culture medium and a preparation method thereof. The composition comprises the following components in parts by weight: 50-100 parts of mushroom dregs, 20-30 parts of wormcast and 3-8 parts of associated microbial inoculum. The dendrobium officinale culture substrate disclosed by the invention does not contain bark components, so that the cost for culturing the dendrobium officinale is reduced, and the synthesis of the nutrient components of dendrobium officinale polysaccharide is promoted by adopting soil infinitesimal bacteria and ceruleus chromophoris as associated microorganisms. The dendrobium officinale can be cultured by adopting the culture medium disclosed by the invention, so that the culture method is simple, the yield is high, and the nutrient content is high.
Description
Technical Field
The invention belongs to the technical field of medicinal plant cultivation, and particularly relates to a dendrobium officinale culture medium and a preparation method thereof.
Background
The Dendrobium officinale mainly contains polysaccharides, alcohol-soluble extracts, alkaloids, amino acids, trace elements and the like, and has the effects of resisting oxidation, resisting fatigue, reducing blood sugar, improving immunity and the like. The stem of Dendrobium officinale has the highest polysaccharide content in roots, stems and leaves, and is the main medicinal part of Dendrobium officinale.
The domestication and cultivation of the dendrobium officinale in 1992 makes a major breakthrough, the planting technology of the dendrobium officinale is well popularized nationwide, and the dendrobium officinale planting industry is developed in more than ten provinces currently nationwide. The existing substrate for cultivating the dendrobium officinale generally adopts a bark cultivation substrate, nutrients such as moss and wormcast are supplemented, a large piece of bark is paved on the bottom layer, nutrients such as small pieces of bark with the moss and the wormcast are added on the upper layer, the natural growth environment of the dendrobium officinale is simulated, the bark needs to be subjected to sterilization treatment before use, the treatment cost is high, the bark is easily infected with harmful bacteria in the cultivation process, and a large amount of pesticides need to be applied to keep the healthy growth of the dendrobium officinale, so that the problem of high pesticide residue of finished products of the dendrobium officinale is caused.
In a natural environment, the dendrobium officinale grows together with various associated beneficial bacteria, the beneficial bacteria promote the generation of nutrient substances of the dendrobium officinale, and the associated beneficial bacteria are lost under artificial culture conditions, so that the artificial culture of the dendrobium officinale has low nutritive value, and the search for proper associated beneficial bacteria to be symbiotic with the dendrobium officinale is a key for culturing the dendrobium officinale with high nutritive value.
Disclosure of Invention
The invention aims to provide a dendrobium officinale culture medium and a preparation method thereof.
A dendrobium officinale culture medium is composed of the following components in parts by weight: 50-100 parts of mushroom dregs, 20-30 parts of wormcast and 3-8 parts of associated microbial inoculum.
The associated microbial agent is a small unicellular microbial agent, and the concentration of live bacteria in the microbial agent is 1-3 multiplied by 108cfu/mL。
The Micromonospora is soil Micromonospora and/or Micromonospora purpurea.
A preparation method of a dendrobium officinale culture medium comprises the following steps:
(1) mixing the mushroom dregs and wormcast uniformly, compressing into sheets, then sterilizing in a sterilizing pot at high temperature for 10-20min, drying in the sun, and controlling the water content to be 15-20%;
(2) selecting small single cell strain, inoculating to liquid culture medium to culture small single cell strain until viable bacteria concentration is 1-3 × 108cfu/mL, spraying the cfu/mL into the mixture prepared in the step (1), packaging and preparing.
The elevated temperature is 115 ℃ or 121 ℃.
The mixture is pressed into a sheet shape in the step (1), the area is 1-2 square centimeters, and the thickness is 2-4 mm.
The liquid culture medium comprises the following components: glucose 9.5 g/L; k2HPO40.8g/L；KH2PO40.2g/L； NH4Cl1.8g/L；CaCl20.2g/L；FeSO4·7H2O 0.08g/L；ZnSO40.001g/L； MgSO4·7H2O 0.05g/L；NaCl 1.2g/L；CuSO40.01g/L；(NH4)6Mo4O24·4H2O 0.001 g/L；VB20.00003 g/L; nicotinic acid 0.00003 g/L; 5g/L of beef extract; 1000mL of water; the initial pH of the liquid medium was adjusted to 7.5-8.0.
The invention has the beneficial effects that: the dendrobium officinale culture substrate disclosed by the invention does not contain bark components, so that the cost for culturing the dendrobium officinale is reduced, and the synthesis of the nutrient components of dendrobium officinale polysaccharide is promoted by adopting soil minimal monad and/or cervus purpureus chromogenic micromod as associated microorganisms. The dendrobium officinale can be cultured by adopting the culture medium disclosed by the invention, so that the culture method is simple, the yield is high, and the nutrient content is high.
Detailed Description
The present invention is further illustrated by the following specific examples.
In the following examples, the test materials selected were as follows:
dendrobium officinale C13 strain, gifted by agriculture and forestry university in Zhejiang.
Soil minimal single cell bacterium (Pusillimmonas soli) purchased from China general microbiological culture Collection center with the preservation number of CGMCC 1.10919.
Micromonospora purpureochromyenes (Micromonospora purpurogens) purchased from China general microbiological culture Collection center with the collection number CGMCC 4.6585.
Component configuration of a small unicellular bacterium liquid culture medium: glucose 9.5 g/L; k2HPO40.8g/L； KH2PO40.2g/L；NH4Cl 1.8g/L；CaCl20.2g/L；FeSO4·7H2O 0.08g/L；ZnSO40.001 g/L；MgSO4·7H2O0.05g/L；NaCl 1.2g/L；CuSO40.01g/L；(NH4)6Mo4O24·4H2O 0.001g/L；VB20.00003 g/L; nicotinic acid 0.00003 g/L; 5g/L of beef extract; 1000mL of water; the initial pH of the liquid medium was adjusted to 7.8.
Example 1
A dendrobium officinale culture medium comprises the following components: 80kg of mushroom dregs, 25kg of wormcast and 5L of soil minimal unicellular bacteria agent, wherein the concentration of viable bacteria in the agent is 2 multiplied by 108cfu/mL。
The preparation method of the dendrobium officinale culture medium comprises the following steps:
(1) mixing the mushroom dregs and wormcast uniformly, compressing into sheets, sterilizing in a sterilizing pot at 115 ℃ for 15min, and drying in the sun, wherein the water content is controlled to be 18%;
(2) selecting soil minimal single cell strain, inoculating to liquid culture medium to culture soil minimal single cell strain until viable bacteria concentration is 2 × 108cfu/mL, spraying the cfu/mL into the mixture prepared in the step (1), packaging and preparing.
Example 2
A dendrobium officinale culture medium is composed of the following components in parts by weight: 60kg of fungus dregs, 28kg of wormcast and 4L of rhodochrous producing small unicellular fungus agent, wherein the concentration of viable bacteria in the fungus agent is 1 multiplied by 108cfu/mL。
The preparation method of the dendrobium officinale culture medium comprises the following steps:
(1) mixing the mushroom dregs and wormcast uniformly, compressing into sheets, sterilizing in a sterilizing pot at 115 ℃ for 12min, and drying in the sun, wherein the water content is controlled to be 19%;
(2) selecting Micromonospora purpurea strain, inoculating to liquid culture medium, and culturing Micromonospora purpurea until the concentration of viable bacteria is 1 × 108cfu/mL, spraying the cfu/mL into the mixture prepared in the step (1), packaging and preparing.
Example 3
A dendrobium officinale culture medium is composed of the following components in parts by weight: 90kg of fungus residues, 26kg of wormcast and 7L of soil minimal unicellular bacterium agent, wherein the concentration of viable bacteria in the bacterium agent is 1.5 multiplied by 108 cfu/mL.
The preparation method of the dendrobium officinale culture medium comprises the following steps:
(1) mixing the mushroom dregs and wormcast uniformly, compressing into sheets, then sterilizing in a sterilizing pot at 121 ℃ for 20min, and drying in the sun, wherein the water content is controlled to be 15%;
(2) selecting soil minimal single cell strain, inoculating to liquid culture medium to culture soil minimal single cell strain until viable bacteria concentration is 1.5 × 108cfu/mL, spraying the cfu/mL into the mixture prepared in the step (1), packaging and preparing.
Comparative example 1
A culture medium for Dendrobium officinale comprisesThe components are as follows: 80kg of pine bark, 25kg of wormcast and 5L of soil minimal unicellular bacteria agent, wherein the concentration of viable bacteria in the agent is 2 multiplied by 108cfu/mL。
The preparation method of the dendrobium officinale culture medium comprises the following steps:
(1) mixing the mushroom dregs and wormcast uniformly, compressing into sheets, sterilizing in a sterilizing pot at 115 ℃ for 15min, and drying in the sun, wherein the water content is controlled to be 18%;
(2) drying wormcast in the sun, grinding into powder, sieving with a 80-mesh sieve, and mixing into the mushroom dreg strips prepared in the step (1);
(3) selecting soil minimal single cell strain, inoculating to liquid culture medium to culture soil minimal single cell strain until viable bacteria concentration is 2 × 108cfu/mL, spraying into the mixture prepared in the step (2), packaging and preparing.
Comparative example 2
A dendrobium officinale culture medium comprises the following components: 80kg of mushroom dregs and 25kg of wormcast.
The preparation method of the dendrobium officinale culture medium comprises the following steps:
(1) mixing the mushroom dregs and wormcast uniformly, compressing into sheets, sterilizing in a sterilizing pot at 115 ℃ for 15min, and drying in the sun, wherein the water content is controlled to be 18%;
(2) drying wormcast in the sun, grinding into powder, sieving with a 80-mesh sieve, mixing with the mushroom residue strips prepared in the step (1), and packaging to obtain the finished product.
Experimental example 1:
the dendrobium officinale culture mediums prepared in the embodiments 1-3 and the control groups 1-2 are subjected to physical property detection, and the detection indexes comprise volume weight (g/cm)3) Total porosity (%) and electrical conductivity (uS/cm), the results are shown in table 1:
TABLE 1
Note: represents P <0.05 compared to control example 1 group.
As can be seen from Table 1, the volume weight of the dendrobium officinale culture medium in each experimental group is not obviously different, the water retention porosity of examples 1-3 and comparative example 2 is obviously higher than that of comparative example 1, the main reason is that the main material added in the comparative example 1 is pine bark, the fungus residue added in other examples is superior to that of the pine bark, and the conductivity of examples 1-3 and comparative example 2 is also obviously higher than that of comparative example 1, so that the fungus residue serving as the culture medium contains more inorganic salt and can provide more inorganic nutrient substances.
Experimental example 2:
2kg of the dendrobium officinale culture medium prepared in the example 1-3 and the comparative example 1-2 is filled in each flowerpot, the dendrobium officinale C13 strain is selected, seedlings with the same weight are transplanted, the color of the seedling leaves is obviously darker, the leaves are thickened, and more than 5 leaves grow on the seedlings; 3-4 internodes are arranged in the stem, 3-5 rhizomes are arranged in the root, watering is carried out once every 3 days, the culture temperature is about 22 ℃, the humidity is controlled to be 65% -75%, harvesting is carried out after 1 year of culture, weighing is carried out, each group of experiments are repeated for 3 times, the average value is taken, and the weight of each potted dendrobium officinale product in each experimental group is shown in table 2:
TABLE 2
Note: represents P <0.05 compared to control example 1 group.
As can be seen from table 2, when pine bark is used to replace the fungus residue of the present invention (comparative example 1), the yield of finished dendrobium officinale is significantly reduced, and it is proved that the yield of dendrobium officinale can be increased by the associated microorganisms, and the associated microorganisms inhibit the growth of harmful bacteria and secrete nutrients to promote the growth of dendrobium officinale in the symbiotic process with dendrobium officinale.
Collecting fresh Dendrobium officinale products of each experimental group, respectively removing leaves of Dendrobium officinale, cleaning, cutting stems into slices, drying in a drying oven at 60 ℃ to constant weight, crushing by a high-speed universal crusher, sieving by a 60-mesh sieve, and placing in a dryer for later use.
1mL of glucose control solutions with concentrations of 5.82. mu.g/m L, 78. mu.g/m L, 98. mu.g/m L, 116.2. mu.g/m L, 135.8. mu.g/m L, 156.2. mu.g/m L and 198. mu.g/m L were weighed, 1mL of a 5% phenol solution was added precisely, and the mixture was shaken well. A blank solution was prepared with ultrapure water and the absorbance was measured at a wavelength of 487nm in an ultraviolet spectrophotometer.
Taking the absorbance as a vertical coordinate and the glucose concentration as a horizontal coordinate, drawing a standard curve, and obtaining a linear regression equation through statistical analysis, wherein the linear regression equation is as follows: y11138 x +0.0509, R20.9996(n 7). The results show that: the linear relation of the grape concentration is good in the range of 5.82 mu g/m L-192 mu g/m L.
Respectively taking 1g of dendrobium officinale powder of each experimental group, adding 200m L of water, carrying out reflux extraction for 2 hours, cooling, transferring the extracting solution into a 250m L measuring flask, washing the container with a proper amount of water in a plurality of times, transferring the washing solution into the same measuring flask, adding water to the scale, and shaking up. Precisely sucking the subsequent filtrate 2m L, placing into a 15m L centrifuge tube, adding anhydrous ethanol 10m L, shaking up, refrigerating for 1 hour, taking out, centrifuging for 25 minutes at 4000 rpm, discarding the supernatant, filtering if necessary, adding 80% ethanol into the precipitate, washing for 2 times, 8ml each time, centrifuging, discarding the supernatant, adding hot water to dissolve the precipitate, transferring into a 25m L measuring flask, cooling, diluting with water to a scale, and shaking up to obtain the sample treatment solution. Measuring the absorbance of the sample treatment solution, performing parallel measurement for 3 times, taking an average value, converting the average value into polysaccharide content according to a standard curve, and obtaining the measurement result shown in table 3:
TABLE 3
Note: represents P <0.05 compared to control example 1 group.
As can be seen from Table 3, the content of polysaccharide in the medium without the addition of the companion microorganism (comparative example 2) is significantly lower than that in other groups, which proves that the companion microorganism has a key effect on the synthesis of dendrobium officinale polysaccharide.
Claims (5)
1. The dendrobium officinale culture medium is characterized by comprising the following components in parts by weight: 50-100 parts of mushroom dregs, 20-30 parts of wormcast and 3-8 parts of associated microbial agent;
the associated microbial agent is soil infinitesimal bacterium or rhodochrous microcystis purpurea, and live bacteria in the microbial agentThe concentration is 1-3 × 108cfu/mL。
2. A preparation method of a dendrobium officinale culture medium is characterized by comprising the following steps:
(1) mixing the mushroom dregs and wormcast uniformly, compressing into sheets, then sterilizing in a sterilizing pot at high temperature for 10-20min, drying in the sun, and controlling the water content to be 15-20%;
(2) selecting small single cell strain, inoculating to liquid culture medium to culture small single cell strain until viable bacteria concentration is 1-3 × 108cfu/mL, spraying the cfu/mL into the mixture prepared in the step (1), packaging and preparing.
3. The method for preparing the dendrobium officinale culture medium according to claim 2, wherein the high temperature is 115 ℃ or 121 ℃.
4. The method for preparing the dendrobium officinale culture substrate according to claim 2, wherein the dendrobium officinale culture substrate obtained in the step (1) is pressed into a sheet shape, the area size is 1-2 square centimeters, and the thickness is 2-4 mm.
5. The preparation method of the dendrobium officinale culture medium according to claim 2, wherein the liquid culture medium comprises the following components: glucose 9.5 g/L; k2HPO40.8g/L；KH2PO40.2g/L；NH4Cl 1.8g/L；CaCl20.2g/L；FeSO4·7H2O 0.08g/L；ZnSO40.001g/L；MgSO4·7H2O 0.05g/L；NaCl 1.2g/L；CuSO40.01g/L；(NH4)6Mo4O24·4H2O 0.001g/L；VB20.00003 g/L; nicotinic acid 0.00003 g/L; 5g/L of beef extract; 1000mL of water; the initial pH of the liquid medium was adjusted to 7.5-8.0.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910280356.8A CN109819868B (en) | 2019-04-09 | 2019-04-09 | Dendrobium officinale culture medium and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910280356.8A CN109819868B (en) | 2019-04-09 | 2019-04-09 | Dendrobium officinale culture medium and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109819868A CN109819868A (en) | 2019-05-31 |
| CN109819868B true CN109819868B (en) | 2020-11-03 |
Family
ID=66874239
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910280356.8A Active CN109819868B (en) | 2019-04-09 | 2019-04-09 | Dendrobium officinale culture medium and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109819868B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110156521A (en) * | 2019-06-14 | 2019-08-23 | 中国热带农业科学院环境与植物保护研究所 | A kind of culture medium and preparation method thereof for cultivating the bletilla striata |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2614101B2 (en) * | 1988-01-16 | 1997-05-28 | 栃木県 | Novel microorganism and method for controlling plant diseases using the same |
| CN103524158B (en) * | 2013-10-10 | 2015-09-16 | 贺州市农村技术发展中心 | Herba Dendrobii planting matrix preparation method |
| CN105112309A (en) * | 2015-09-25 | 2015-12-02 | 江苏农林职业技术学院 | Umbelopsis nana and application thereof in promoting growth of Dendrobium officinale |
| CN106305193A (en) * | 2016-08-24 | 2017-01-11 | 福建农业职业技术学院 | Planting frame and cultivation technology for cultivating dendrobium officinale by planting frame |
| CN106497838A (en) * | 2016-10-28 | 2017-03-15 | 云南山里红生物科技有限公司 | A kind of composite bacteria agent capable of Herba Dendrobii |
| CN108371090A (en) * | 2016-11-20 | 2018-08-07 | 石佳媛 | Application of the mycorrhizal fungi in dendrobe cultivation |
-
2019
- 2019-04-09 CN CN201910280356.8A patent/CN109819868B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN109819868A (en) | 2019-05-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101491195B (en) | Phlebopus portentosus cultivation method | |
| CN104322273B (en) | Technique planted by the artificial bag of Phellinus igniarius (L. ex Fr.) Quel. | |
| CN106490003B (en) | Inhibition and its extracting method of the celery root exudates concentrate to ralstonia solanacearum of tomato | |
| KR20200076934A (en) | Method for obtaining mass proliferation of Cordyceps sinensis and mass culture method using it | |
| CN103103152B (en) | Corallococcus coralloides and application thereof | |
| CN108477232B (en) | Plant source thinning and removing agent for fruit trees and preparation method thereof | |
| JP7393824B2 (en) | Organic cultivation method of red cedar | |
| Koleva Gudeva et al. | Influence of different auxins on rooting of rosemary, sage and elderberry | |
| CN112359084A (en) | Preparation and prevention and treatment application of mycoprotein in bacillus subtilis fermentation product | |
| CN102061330B (en) | Method for identifying pathogenicity of sesame stem blight and blast pathogenic bacteria | |
| CN109819868B (en) | Dendrobium officinale culture medium and preparation method thereof | |
| CN111052997B (en) | Preparation and application method of biological stimulin for improving strawberry continuous cropping obstacle resistance | |
| CN105961441B (en) | The application of bacillus JC65 and its volatile materials in plant growth-promoting | |
| CN101781630B (en) | Root nodule azotobacter strain RY1 bacterial strain and application thereof | |
| CN114774300B (en) | Pseudomonas koraiensis and application thereof | |
| Mbong et al. | Influence of cropping system on the incidence and severity of leaf spot disease of Telfairia occidentalis Hook f. caused by Phoma sorghina | |
| CN113615538A (en) | Substrate applied to Hangzhou white chrysanthemum cultivation and preparation method | |
| CN112106789A (en) | Application of delphinium ethyl acetate extract in preventing and treating diamond back moth | |
| CN111304097A (en) | Wild pleurotus ferulae strain separation and purification method | |
| KR101821349B1 (en) | Method for mass production of Poria cocos using a stump inoculation | |
| KR100203516B1 (en) | The method of cultivating orchid | |
| CN103333852A (en) | Production method of rhizopus oryzae microbial inoculum and use and using method of rhizopus oryzae microbial inoculum | |
| CN117535201B (en) | Bacillus bailii HM-6 and application thereof | |
| CN109988736B (en) | Pine endophytic bacterium FBS135 and application thereof | |
| CN115369046B (en) | Trichoderma harzianum for preventing and treating various diseases of vegetables and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |