CN113229145A

CN113229145A - Technology for carrying out spray breeding on dendrobium huoshanense cells

Info

Publication number: CN113229145A
Application number: CN202110456996.7A
Authority: CN
Inventors: 陈大卫
Original assignee: Individual
Current assignee: Individual
Priority date: 2021-04-27
Filing date: 2021-04-27
Publication date: 2021-08-10

Abstract

The invention discloses a technology for spray breeding of dendrobium huoshanense cells, which comprises the following steps of selecting dendrobium huoshanense branches with high plant height and thick stems, washing and disinfecting the dendrobium huoshanense branches, dissolving small dendrobium huoshanense branches by using cellulase, vibrating by a shaking table and centrifuging by a centrifuge, filtering to obtain dendrobium huoshanense cells, and finally spraying the dendrobium huoshanense cells in a culture medium for breeding. The technique of dendrobium huoshanense cell spraying breeding, through add cellulase and pectinase when the isolated cell, can break away from out in the dendrobium huoshanense cell follow dendrobium huoshanense branch fast, and through adding coconut juice and potato juice liquid in the culture medium, coconut juice can promote cell division, potato juice liquid can cushion the PH of culture medium, be favorable to obtaining the plant of being strong, carry out the reproduction of dendrobium huoshanense through the cell, can obtain a large amount of pure kind dendrobium huoshanense young plants fast, after handling through the wild, can alleviate the demand of market to dendrobium huoshanense.

Description

Technology for carrying out spray breeding on dendrobium huoshanense cells

Technical Field

The invention relates to the technical field of agriculture, in particular to a technology for spray breeding of dendrobium huoshanense cells.

Background

Forty medicinal plants are present in 76 dendrobium plants in China. The pharmacology of the medicine is mainly to strengthen yin and benefit body fluid, tonify kidney and benefit strength, improve eyesight and prolong life. However, due to the fact that a large number of dendrobium are picked, the dendrobium grows slowly, the growth period is long, wild dendrobium is quite rare, and good quality is difficult to achieve if any.

Dendrobium huoshanense, also known as Dendrobium huoshanense, is the top grade of medicinal Dendrobium nobile. The dendrobium huoshanense has extremely strict requirements on growth conditions, and wild dendrobium plants are endangered to be extinct due to long-term artificial damage and excessive mining and pulling.

In order to relieve the situation that wild dendrobium is endangered and extincted, the wild dendrobium seedlings are cultivated artificially and then are subjected to wild treatment, but the dendrobium seedlings have low self-reproduction capacity and cannot meet the requirement of large-area wild.

Disclosure of Invention

The invention aims to solve the defects in the prior art and provides a technology for spraying and breeding dendrobium huoshanense cells.

In order to achieve the purpose, the invention adopts the following technical scheme:

a technology for carrying out spray breeding on dendrobium huoshanense cells comprises the following steps:

collecting dendrobium huoshanense branches, and selecting the branches with good appearance indexes such as plant height, stem thickness and the like from the collected dendrobium huoshanense branches;

placing the selected dendrobe branches in a flowing water tank, standing and washing for 30min, and washing off soil and impurities stained on the dendrobe branches;

step three, immersing the dendrobium candidum branches obtained after soil and impurities are removed in the step two in a disinfectant for 20-30 min;

step four, washing the dendrobe branches disinfected in the step three by using distilled water, wherein the washing frequency is 3-5 times, and 3min is carried out for each time, so that residual disinfectant on the surfaces of the dendrobe branches can be washed, and the residual disinfectant is avoided;

step five, cutting up the dendrobium huoshanense branches by using a knife and tweezers, putting the dendrobium huoshanense branches into a wide-mouth bottle, adding cellulase into the wide-mouth bottle, putting the wide-mouth bottle into a 900rpm shaking table for treatment for 6 hours, then transferring the wide-mouth bottle into a sterile centrifuge tube, centrifuging the wide-mouth bottle at 1000rpm for 1min, removing supernatant, and collecting dendrobium huoshanense cells;

step six, uniformly spraying the dendrobium huoshanense cells obtained in the step five into 1/2MS culture medium for culture, and placing a culture dish in a sterile laboratory for 60 days to obtain dendrobium huoshanense seedlings;

and step seven, transplanting the dendrobium huoshanense seedlings obtained in the step six into a cultivation pot.

Preferably, the disinfectant in the third step is a 2% sodium hypochlorite solution, and before soaking in the sodium hypochlorite solution, the dendrobe branches are washed with 70% alcohol for 20-30 s.

Preferably, the cell separation procedure in step five is repeated three times and pectinase is added to the jar before the second and third shaking table treatments.

Preferably, in the sixth step, 1/2MS culture medium is added with 10% -20% coconut juice and potato juice.

Preferably, the potato juice is prepared by peeling and sprouting potato, decocting in water for 30min, and filtering to obtain potato juice.

Preferably, in the sixth step, the air humidity of the sterile laboratory is kept at about 50% in the daytime and at about 90% at night.

Preferably, the culture pot in the seventh step is filled with a matrix mixed by peat moss, fern roots, bark blocks and charcoal, meanwhile, tiles and broken bricks are arranged at the bottom of the pot, and in the culture process, 1000 times diluted Huabao No. 1 is sprayed to stems and leaves of young plants every other week.

Drawings

FIG. 1 is an operation flow chart of a technique for spray breeding of Dendrobium huoshanense cells provided by the present invention.

The invention has the following beneficial effects:

1. this technique that dendrobium huoshanense cell carries out spraying breeding, through add cellulase and pectinase when the isolated cell, can break away from out in the dendrobium huoshanense branch with the dendrobium huoshanense cell fast, and through adding coconut juice and potato juice liquid in the culture medium, coconut juice can promote cell division, potato juice liquid can cushion the PH of culture medium, be favorable to obtaining robust plant, carry out the reproduction of dendrobium huoshanense through the cell, can obtain a large amount of pure kind dendrobium huoshanense young plants fast, after handling through the field, can alleviate the demand of market to dendrobium huoshanense.

2. This technique of dendrobium huoshanense cell spraying breeding, through cultivateing under sterile environment, and with the humidity control of aseptic laboratory in the humidity range that is fit for stem of noble dendrobium and grows, spray diluted huabao No. 1 to the stem leaf of young plant every week, be favorable to the rooting of young plant.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments.

In the description of the present invention, it is to be understood that the terms "upper", "lower", "front", "rear", "left", "right", "top", "bottom", "inner", "outer", and the like, indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, are merely for convenience in describing the present invention and simplifying the description, and do not indicate or imply that the device or element being referred to must have a particular orientation, be constructed and operated in a particular orientation, and thus, should not be construed as limiting the present invention.

The embodiment for spray breeding of dendrobium huoshanense cells comprises the following steps:

step three, immersing the dendrobium candidum branches obtained after soil and impurities are removed in the step two in a 2% sodium hypochlorite solution for 20-30 min, and washing the dendrobium candidum branches for 20-30 s by using 70% alcohol before immersing in the sodium hypochlorite solution;

step four, washing the dendrobium huoshanense branches sterilized in the step three by using distilled water, wherein the washing frequency is 3-5, and 3min is carried out for each washing, so that residual sterilizing agents on the surfaces of the dendrobium huoshanense branches can be washed, and agent residues are avoided;

step five, cutting the dendrobium huoshanense branches into pieces by using a knife and tweezers, putting the dendrobium huoshanense branches into a wide-mouth bottle, adding cellulase into the wide-mouth bottle, placing the wide-mouth bottle on a 900rpm shaking table for treatment for 6 hours, enabling the cellulase to fully dissolve fibers in the dendrobium huoshanense branches, then transferring the dendrobium huoshanense branches into an aseptic centrifuge tube, centrifuging the centrifuge tube at 1000rpm for 1min, removing supernatant, repeating the operation for three times, adding pectinase into the wide-mouth bottle before the second shaking table treatment and the third shaking table treatment, enabling the pectinase to accelerate the dispersion effect among cells, quickly separating dendrobium huoshanense cells from the dendrobium huoshanense branches by adding the cellulase and the pectinase during cell separation, then filtering dendrobium huoshanense cell suspension by using a stainless steel screen, removing large cell clusters and impurities, and collecting small cell clusters with a certain volume;

step six, uniformly spraying the dendrobium candidum cells obtained in the step five into an 1/2MS culture medium for culture, adding 10-20% of coconut juice and potato juice into a 1/2MS culture medium, wherein the coconut juice can promote cell division, the potato juice has a large buffering effect on PH, a strong plant can be obtained after adding, placing a culture dish in an aseptic laboratory for 60 days, controlling the air humidity of the aseptic laboratory at about 50% in the daytime and controlling the air humidity at about 90% at night, and the dendrobium candidum plants can grow favorably, so that healthy dendrobium candidum seedlings can be obtained;

and seventhly, transplanting the dendrobium huoshanense seedlings obtained in the sixth step into a cultivation pot, filling a matrix mixed by peat moss, fern roots, bark blocks and charcoal into the cultivation pot, simultaneously laying tiles and broken bricks at the bottom of the pot, spraying and diluting 1000 times of Huabao No. 1 to stems and leaves of young plants every other week in the cultivation process, and spraying and diluting Huabao No. 1 periodically to the stems and leaves of the young plants, so that the young plants of the dendrobium huoshanense can take roots conveniently.

In the embodiment, the small cell clusters obtained in the fifth step are respectively smeared in 1/2MS culture medium without coconut juice and potato juice and 1/2MS culture medium with 10% -20% coconut juice and potato juice, the culture is carried out for 60 days under the same environment, and the branch circumference and the number of branches of dendrobium candidum seedlings cultured by two groups of different culture media are compared, so that dendrobium candidum seedlings cultured in 1/2MS culture medium added with 10% -20% coconut juice and potato juice are obviously stronger than the dendrobium candidum seedlings cultured in 1/2MS culture medium without coconut juice and potato juice.

Table one is a table comparing the stem circumference and the number of branches of the dendrobe seedlings obtained from two different culture media.

Watch 1

In the embodiment, the dendrobium candidum seedlings obtained by culturing in the sixth step by using 1/2MS culture medium containing 10-20% of coconut juice and potato juice are placed under different air humidities by using the same substrate for culturing for 60 days (the humidity is controlled to be about 50% in the daytime and about 90% in the nighttime, the humidity is not controlled, and the air humidity is kept between 40-50%), the humidity of the dendrobium candidum growth environment is controlled under the same nutrition supply by comparing the increase of the stem circumference and the number of branches of the dendrobium candidum seedlings, and the growth of dendrobium candidum can be accelerated.

And the second table is a comparison table of the stem circumference and the number growth amount of the branches of the dendrobium candidum seedlings under two groups of different humidity environments.

Watch two

According to the dendrobium huoshanense seedling cultivation method, dendrobium huoshanense seedlings of pure varieties can be obtained quickly by extracting dendrobium huoshanense cells and breeding by using the dendrobium huoshanense cells, in the cultivation process, 10% -20% of coconut juice and potato juice are added into a culture medium, dendrobium huoshanense seedlings can be obtained more robustly, the growth of dendrobium huoshanense seedlings can be promoted by controlling the environment humidity during transplanting (the air humidity in the daytime is controlled to be about 50%, and the air humidity at night is controlled to be about 90%), the cells are used for breeding, a large number of seedlings can be obtained in a short time, and the demand of markets for dendrobium huoshanense can be relieved after wild cultivation.

The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.

Claims

1. A technology for carrying out spray breeding on dendrobium huoshanense cells is characterized in that: the method comprises the following steps:

2. The technology for spray breeding of dendrobium huoshanense cells as claimed in claim 1, wherein: and in the third step, the disinfectant is a 2% sodium hypochlorite solution, and before the disinfectant is soaked in the sodium hypochlorite solution, the dendrobe branches are washed for 20-30 s by using 70% alcohol.

3. The technology for spray breeding of dendrobium huoshanense cells as claimed in claim 1, wherein: the cell separation operation in step five can be repeated three times, and pectinase is added into the wide-mouth bottle before the second shaking table treatment and the third shaking table treatment.

4. The technology for spray breeding of dendrobium huoshanense cells as claimed in claim 1, wherein: in the sixth step, the 1/2MS culture medium is added with 10 to 20 percent of coconut juice and potato juice.

5. The technology of claim 4, wherein the technology comprises the following steps: the potato juice is prepared by peeling and bud removing potato, decocting in water for 30min, and filtering.

6. The technology for spray breeding of dendrobium huoshanense cells as claimed in claim 1, wherein: and step six, keeping the air humidity of the sterile laboratory at about 50% in the daytime and about 90% at night.

7. The technology for spray breeding of dendrobium huoshanense cells as claimed in claim 1, wherein: and seventhly, filling a matrix mixed by peat moss, fern roots, bark blocks and charcoal in the cultivation pot, simultaneously laying tiles and broken bricks at the bottom of the pot, and spraying and diluting 1000 times of Huabao No. 1 to the stem and leaf of the young plant every other week in the cultivation process.