CN109819868B - A kind of Dendrobium officinale cultivation substrate and preparation method thereof - Google Patents

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

A kind of Dendrobium officinale cultivation substrate and preparation method thereof

technical field

The invention belongs to the technical field of medicinal plant cultivation, and particularly relates to a Dendrobium officinale cultivation substrate and a preparation method thereof.

Background technique

Dendrobium officinale mainly contains polysaccharides, alcohol-soluble extracts, alkaloids, amino acids, trace elements, etc. The roots, stems and leaves of Dendrobium candidum have the highest polysaccharide content and are the main medicinal parts of Dendrobium candidum.

In 1992, a major breakthrough was made in the domestication and cultivation of Dendrobium officinale, and its planting technology has been well promoted nationwide. Now more than a dozen provinces across the country have developed Dendrobium candidum planting. Existing substrates for cultivating Dendrobium officinale generally use bark cultivation substrates, supplemented with nutrients such as moss and vermicompost, the bottom layer is covered with large pieces of bark, and the upper layer is small pieces of bark plus moss, vermicompost and other nutrients, imitating Dendrobium candidum. In the natural growth environment, the bark needs to be sterilized before use, and the treatment cost is high. During the cultivation process, the bark is easily contaminated with harmful bacteria. The problem of high pesticide residues, if no pesticides are applied, Dendrobium officinale will be infected with harmful bacteria, the products will be different, and the content of nutrients such as polysaccharides will be low.

In the natural environment, Dendrobium officinale grows with a variety of associated beneficial bacteria. The beneficial bacteria promote the formation of nutrients in Dendrobium officinale, and the associated beneficial bacteria are lost under artificial cultivation conditions. Therefore, artificially cultivated Dendrobium officinale has low nutritional value. The symbiosis of the associated beneficial bacteria and Dendrobium officinale is the key to cultivating high nutritional value of Dendrobium officinale.

SUMMARY OF THE INVENTION

The purpose of the present invention is to provide a Dendrobium officinale cultivation substrate and a preparation method thereof.

A Dendrobium officinale cultivation substrate is composed of the following components in parts by weight: 50-100 parts of fungal residue, 20-30 parts of vermicompost, and 3-8 parts of an associated microbial inoculant.

The accompanying microbial inoculum is a Micromonas inoculum, and the concentration of viable bacteria in the inoculum is 1-3×10 ⁸ cfu/mL.

The Micromonas is Micromonas soil micromonas and/or Micromonas cyanobacteria.

A preparation method of a Dendrobium officinale cultivation substrate is carried out according to the following steps:

(1) Mix the fungus residue and vermicompost evenly, press-mold into a sheet, then send it into a sterilizing pot for sterilization at high temperature for 10-20min, and dry it in the sun to control the moisture content at 15-20%;

(2) Pick Micromonas strain, inoculate it into a liquid medium to cultivate Micromonas until the concentration of viable bacteria is 1-3×10 ⁸ cfu/mL, and spray it onto the mixture prepared in step (1). in, packaged, made.

The high temperature is 115°C or 121°C.

Step (1) is compression-molded into a sheet, with an area of 1-2 square centimeters and a thickness of 2-4 mm.

The components of the liquid medium are: glucose 9.5g/L; K ₂ HPO ₄ 0.8g/L; KH ₂ PO ₄ 0.2g/L; NH ₄ Cl 1.8g/L; CaCl ₂ 0.2g/L; FeSO ₄ 7H ₂ O 0.08g/L; ZnSO ₄ 0.001g/L; MgSO ₄ 7H ₂ O 0.05g/L; NaCl 1.2g/L; CuSO ₄ 0.01g/L; (NH ₄ ) ₆ Mo ₄ O ₂₄ · 4H ₂ O 0.001 g/L; V _B2 0.00003 g/L; niacin 0.00003 g/L; beef extract 5 g/L; water 1000 mL; the initial pH value of the liquid medium was adjusted to 7.5-8.0.

Beneficial effects of the present invention: the Dendrobium officinale cultivation substrate of the present invention does not contain bark components, reduces the cost of cultivating Dendrobium officinale, and adopts Soil Micromonas and/or Micromonas cyanobacteria as accompanying microorganisms, Promote the synthesis of Dendrobium officinale polysaccharide nutrients. Using the cultivation substrate of the invention to cultivate Dendrobium officinale has the advantages of simple cultivation method, high yield and high nutrient content.

Detailed ways

The present invention will be further described below with reference to specific embodiments.

In the following examples, the selected test materials are as follows:

Dendrobium officinale C13 strain, donated by Zhejiang Agriculture and Forestry University.

Pusillimonas soli was purchased from the China General Microorganism Culture Collection and Management Center, with the deposit number CGMCC 1.10919.

Micromonospora purpureochromogenes was purchased from China General Microorganism Culture Collection and Management Center, with the deposit number CGMCC 4.6585.

Component configuration of Micromonas liquid medium: glucose 9.5g/L; K ₂ HPO ₄ 0.8g/L; KH ₂ PO ₄ 0.2g/L; NH ₄ Cl 1.8g/L; CaCl ₂ 0.2g/L ; FeSO ₄ ·7H ₂ O 0.08g/L; ZnSO ₄ 0.001 g/L; MgSO ₄ ·7H ₂ O 0.05g/L; NaCl 1.2g/L; CuSO ₄ 0.01g/L; (NH ₄ ) ₆ Mo ₄ O ₂₄ ·4H ₂ O 0.001 g/L; V _B2 0.00003 g/L; niacin 0.00003 g/L; beef extract 5 g/L; water 1000 mL; the initial pH value of the liquid medium was adjusted to 7.8.

Example 1

A Dendrobium officinale cultivation substrate is composed of the following components: 80 kg of fungus residue, 25 kg of vermicompost, 5 L of a soil micromonas bacteria agent, and the concentration of viable bacteria in the bacteria agent is 2×10 ⁸ cfu/mL.

The preparation method of above-mentioned Dendrobium officinale cultivation substrate is carried out according to the following steps:

(1) Mix the fungus residue and vermicompost evenly, press-mold it into a sheet, then send it to a sterilizing pot for sterilization at 115°C for 15 minutes, and dry it in the sun to control the moisture content at 18%;

(2) Picking the strain of Micromonas Soil, inoculating it into the liquid medium to cultivate the Soil Minimonas until the concentration of viable bacteria is 2×10 ⁸ cfu/mL, and spraying it on the prepared micromonas in step (1). Mixed, packaged, made.

Example 2

A kind of Dendrobium officinale cultivation substrate is made up of the following components by weight: 60kg of fungus residue, 28kg of vermicompost, 4L of Micromonas chromogenes inoculum, and the concentration of viable bacteria in the inoculum is 1×10 ⁸ cfu/mL.

The preparation method of above-mentioned Dendrobium officinale cultivation substrate is carried out according to the following steps:

(1) Mix the fungus residue and the vermicompost evenly, press-mold it into a sheet, then send it into a sterilizer for sterilization at 115°C for 12 minutes, and dry it in the sun to control the moisture content at 19%;

(2) pick the Micromonas chromogens strain, inoculate it into the liquid medium to cultivate Micromonas chromogens, to the concentration of viable bacteria is 1 × 10 ⁸ cfu/mL, spray to step ( 1) The prepared mixture is packaged and made.

Example 3

A Dendrobium officinale cultivation substrate is composed of the following components in parts by weight: 90kg of fungus residue, 26kg of vermicompost, 7L of soil micromonas inoculum, and the concentration of viable bacteria in the inoculum is 1.5×10 cfu/ mL.

The preparation method of above-mentioned Dendrobium officinale cultivation substrate is carried out according to the following steps:

(1) Mix the fungus residue and vermicompost evenly, press-mold it into a sheet, then send it into a sterilizing pot for sterilization at 121°C for 20 minutes, and dry it in the sun to control the moisture content at 15%;

(2) Picking out Micromonas Soil strains, inoculating them into a liquid medium to cultivate Soil Minimonas Soil, until the concentration of viable bacteria is 1.5×10 ⁸ cfu/mL, and spraying on the prepared micromonas in step (1) Mixed, packaged, made.

Comparative Example 1

A Dendrobium officinale cultivation substrate is composed of the following components: 80 kg of pine bark, 25 kg of vermicompost, 5 L of a soil micromonas bacteria agent, and the concentration of viable bacteria in the bacteria agent is 2×10 ⁸ cfu/mL.

The preparation method of above-mentioned Dendrobium officinale cultivation substrate is carried out according to the following steps:

(1) Mix the fungus residue and vermicompost evenly, press-mold it into a sheet, then send it to a sterilizing pot for sterilization at 115°C for 15 minutes, and dry it in the sun to control the moisture content at 18%;

(2) vermicompost is sun-dried, ground into powder, sieved with 80 meshes, and mixed into the fungus residue bar prepared by step (1);

(3) Picking the strain of Micromonas Soil, inoculating it into the liquid medium to cultivate Soil Minimonas until the concentration of viable bacteria is 2 × 10 ⁸ cfu/mL, and spraying it into the prepared solution in step (2). Mixed, packaged, made.

Comparative Example 2

A Dendrobium officinale cultivation substrate is composed of the following components: 80kg of fungus residue and 25kg of vermicompost.

The preparation method of above-mentioned Dendrobium officinale cultivation substrate is carried out according to the following steps:

(1) Mix the fungus residue and vermicompost evenly, press-mold it into a sheet, then send it to a sterilizing pot for sterilization at 115°C for 15 minutes, and dry it in the sun to control the moisture content at 18%;

(2) sun-drying the vermicompost, grinding into powder, passing through an 80-mesh sieve, mixing into the fungus residue bar prepared in step (1), packaging, and making.

Experimental example 1:

The physical properties of the Dendrobium officinale cultivation substrates prepared in Example 1-3 and the control group 1-2 were tested, and the test indicators included bulk density (g/cm ³ ), total porosity (%) and electrical conductivity (uS/cm), and the detection The results are shown in Table 1:

Table 1

Note: * represents P<0.05 compared with control group 1.

As can be seen from Table 1, there is no significant difference in the bulk density of the Dendrobium officinale cultivation substrate in each experimental group, and the water holding porosity of Examples 1-3 and Comparative Example 2 is significantly higher than that of Comparative Example 1, mainly due to the main material added in Comparative Example 1. It is pine bark, and what other embodiments add is fungus slag, the water retention performance of fungus slag is better than pine bark, and the conductivity of Examples 1-3 and Comparative Example 2 is also significantly higher than that of Comparative Example 1, which proves that bacterial slag is used as a culture medium, Contains more inorganic salts, can provide more inorganic nutrients.

Experimental example 2:

In each flowerpot, the Dendrobium officinale cultivation substrate prepared by 2kg of Example 1-3 and Comparative Example 1-2 was prepared, and the Dendrobium officinale C13 strain was selected for use, and the seedlings of the same weight were transplanted. , there are more than 5 leaves; the stem has 3-4 internodes, the root has 3-5, watering every 3 days, the cultivation temperature is about 22 ℃, the humidity is controlled at 65%-75%, the cultivation is 1 Harvest in a year, weigh, repeat 3 times of each test group to get an average value, the weight of each pot of finished Dendrobium officinale in each experimental group is as shown in Table 2:

Table 2

Note: * represents P<0.05 compared with control group 1.

As can be seen from Table 2, adopt pine bark to replace the fungus slag of the present invention (Comparative Example 1), the output of finished product Dendrobium officinale significantly declines, it is proved that the associated microorganism can improve the output of Dendrobium officinale, and the associated microorganism is in the process of symbiosis with Dendrobium officinale. , inhibit the growth of harmful bacteria, and secrete nutrients to promote the growth of Dendrobium officinale.

The fresh products of Dendrobium officinale in each experimental group were harvested, and the Dendrobium officinale leaves were removed and washed respectively. The stems were cut into thin slices and placed in an oven at 60°C to dry to constant weight, pulverized by a high-speed universal pulverizer, passed through a 60-mesh sieve, and placed Reserve in the dryer.

Measure 1 mL of glucose reference solution with concentrations of 5.82 μg/mL, 78 μg/mL, 98 μg/mL, 116.2 μg/mL, 135.8 μg/mL, 156.2 μg/mL, and 198 μg/mL, respectively. Precisely add 1ml of 5% phenol solution and shake well. A blank solution was prepared with ultrapure water, and the absorbance was measured by an ultraviolet spectrophotometer at a wavelength of 487 nm.

Taking the absorbance as the ordinate and the glucose concentration as the abscissa, draw a standard curve, and the linear regression equation obtained by statistical analysis is: Y=11138x+0.0509, R ² =0.9996 (n=7). The results showed that the linear relationship was good in the range of grape concentration from 5.82μg/mL to 192μg/mL.

Take 1 g of Dendrobium officinale powder from each experimental group, add 200 mL of water, reflux for extraction for 2 hours, let cool, transfer the extract to a 250 mL volumetric flask, wash the container with an appropriate amount of water, transfer the lotion to the same volumetric flask, and add water. to mark, shake well. Accurately draw 2 mL of the continued filtrate, put it in a 15 mL centrifuge tube, add 10 mL of absolute ethanol, shake well, refrigerate for 1 hour, take out, centrifuge at 4000 rpm for 25 minutes, discard the supernatant, filter if necessary, and precipitate Add 80% ethanol to wash twice, 8 ml each time, centrifuge, discard the supernatant, dissolve the precipitate in heated water and transfer it to a 25 mL volumetric flask, let cool, dilute to the mark with water, and shake well to obtain the sample treatment solution. Measure the absorbance of the sample treatment solution, measure 3 times in parallel and take the average value, convert it to the polysaccharide content according to the standard curve, and the measurement results are shown in Table 3:

table 3

Note: * represents P<0.05 compared with control group 1.

It can be seen from Table 3 that the polysaccharide content of the medium without associated microorganisms (control example 2) was significantly lower than that of the other groups, proving that the associated microorganisms play a key role in the synthesis of Dendrobium officinale polysaccharide.

Claims (5)

1. a Dendrobium officinale cultivation substrate, is characterized in that, is made up of the component of following parts by weight: 50-100 parts of fungus residues, 20-30 parts of vermicompost, 3-8 parts of associated microbial inoculants; The associated microbial inoculant is Micromonas vermicularis or Micromonas cyanochromogenes, and the concentration of viable bacteria in the inoculum is 1-3×10 ⁸ cfu/mL. 2. a preparation method of Dendrobium officinale cultivation substrate, is characterized in that, carry out according to the following steps: (1) Mix the fungus residue and vermicompost evenly, press-mold into a sheet, then send it into a sterilizing pot for sterilization at high temperature for 10-20min, and dry it in the sun to control the moisture content at 15-20%; (2) Pick Micromonas strain, inoculate it into a liquid medium to cultivate Micromonas until the concentration of viable bacteria is 1-3×10 ⁸ cfu/mL, and spray it onto the mixture prepared in step (1). in, packaged, made. 3 . The preparation method of the Dendrobium officinale cultivation substrate according to claim 2 , wherein the high temperature is 115° C. or 121° C. 4 . 4. the preparation method of the Dendrobium officinale cultivation substrate according to claim 2, is characterized in that, described in step (1), is compression molded into sheet shape, and the area size is 1-2 square centimeters, and the thickness is 2-4mm. 5. the preparation method of the Dendrobium officinale cultivation substrate according to claim 2, is characterized in that, the component of described liquid medium is: glucose 9.5g/L; K ₂ HPO ₄ 0.8g/L; KH ₂ PO ₄ 0.2 g/L; NH ₄ Cl 1.8g/L; CaCl ₂ 0.2g/L; FeSO ₄ ·7H ₂ O 0.08g/L; ZnSO ₄ 0.001g/L; MgSO ₄ ·7H ₂ O 0.05g/L; NaCl 1.2 g/L; CuSO ₄ 0.01g/L; (NH ₄ ) ₆ Mo ₄ O ₂₄ ·4H ₂ O 0.001g/L; V _B2 0.00003g/L; niacin 0.00003g/L; beef extract 5g/L; water 1000mL; adjust the initial pH of the liquid medium to 7.5-8.0.