CN115812604B - Leek regeneration medium and preparation method thereof - Google Patents
Leek regeneration medium and preparation method thereof Download PDFInfo
- Publication number
- CN115812604B CN115812604B CN202310017469.5A CN202310017469A CN115812604B CN 115812604 B CN115812604 B CN 115812604B CN 202310017469 A CN202310017469 A CN 202310017469A CN 115812604 B CN115812604 B CN 115812604B
- Authority
- CN
- China
- Prior art keywords
- chitosan
- alginic acid
- chives
- amphiphilic
- acid
- 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
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
- Y02P60/21—Dinitrogen oxide [N2O], e.g. using aquaponics, hydroponics or efficiency measures
Landscapes
- Agricultural Chemicals And Associated Chemicals (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
The invention discloses a chives regeneration culture medium and a preparation method thereof, belonging to the field of chives culture medium preparation, wherein the preparation raw materials of the chives regeneration culture medium comprise: hydrophobic chitosan, amphiphilic alginic acid, dodecylaminopropionic acid, naphthylacetic acid, absolute ethyl alcohol, indoleacetic acid, calcium chloride solution, agar, sucrose, MS culture medium and 6-benzylaminoadenine. The preparation method comprises the following steps: preparing amphiphilic chitosan-alginic acid, preparing a plant hormone solution, preparing a slow-release particle suspension and preparing a chives regeneration medium. The prepared regeneration culture medium for the allium tuberosum is used for rooting culture of the allium tuberosum in the rooting culture process, and solves the problem that the rooting rate and the survival rate of the allium tuberosum cannot be improved simultaneously in the rooting culture process.
Description
Technical Field
The invention relates to the field of preparation of a chives culture medium, in particular to a chives regeneration culture medium and a preparation method thereof.
Background
The Allium schoenoprasum is perennial herb of Allium of Liliaceae, and is named as Allium schoenoprasum and Allium schoenoprasum. Wild leek resources are very abundant and widely distributed in typical grasslands and forest grassland communities in European and Asian areas. The leeks have a plurality of excellent characteristics and strong adaptability, and can well grow even on drought-barren lands. The distribution range of the Chinese chives is wide, the economic value is high, and the Chinese chives have various values such as eating, medicinal use, feeding, ornamental use and the like: the whole grass can be used as food or flavoring; has antibacterial, antiinflammatory, and anticancer effects, and can be used for preventing and treating hyperlipidemia and thrombosis; the chives contain high levels of protein, vitamins and sugar, and can be used as a high-grade pasture for fattening sheep, horses and camels, and also for controlling infections of parasites in the nasopharynx and intestinal tracts of livestock.
Along with the development of molecular biotechnology, a high-efficiency regeneration system is established by utilizing leaf tissues with rich sources, which is beneficial to realizing large-scale cultivation and mass production of the allium tuberosum. However, although the common tissue culture method can realize the regeneration culture of the chives in a laboratory, due to different hormone effects in the regeneration culture process of the chives, various hormones in a common culture medium act simultaneously, the effect is difficult to control, so that the rooting rate and the survival rate of rooting culture in the culture process cannot be improved simultaneously, and the regeneration culture effect is limited. Therefore, the preparation of the regeneration culture medium for the allium tuberosum can effectively control the slow release of the plant hormone, so as to solve the problem that the rooting rate and the survival rate cannot be improved simultaneously in the rooting culture process.
Disclosure of Invention
In view of the above, the invention aims to provide a regeneration culture medium for chives and a preparation method thereof, which are used for solving the problem that the rooting rate and the survival rate of the chives cannot be improved simultaneously after rooting culture because hormone action is difficult to control in the regeneration culture process of the chives.
The invention solves the technical problems by the following technical means:
the preparation raw materials of the chives regeneration culture medium comprise: hydrophobic chitosan, amphiphilic alginic acid, dodecylaminopropionic acid, naphthylacetic acid, absolute ethyl alcohol, indoleacetic acid, calcium chloride solution, agar, sucrose, MS culture medium and 6-benzylaminoadenine.
Further, the preparation raw materials of the hydrophobic chitosan comprise: chitosan, 1-hydroxybenzotriazole, N-acetyl cysteine, carbodiimide and sodium hydroxide solution.
Further, the preparation raw materials of the amphiphilic alginic acid comprise: alginic acid, cholesterol, dicyclohexylcarbodiimide and 2, 2-dimethylolpropionic acid.
The invention also provides a preparation method of the chives regeneration medium, which comprises the following specific steps:
(1) Preparation of amphiphilic chitosan-alginic acid: adding hydrophobic chitosan into absolute ethyl alcohol for fully dissolving, adding dodecylaminopropionic acid, continuously stirring for 20-30min, and then adding amphiphilic alginic acid, continuously stirring for 1-2h to obtain amphiphilic chitosan-alginic acid for later use;
(2) Preparing a plant hormone solution: fully dissolving naphthalene acetic acid and indole acetic acid in deionized water to obtain a plant hormone solution for later use;
(3) Preparing a slow release particle suspension: pouring amphiphilic chitosan-alginic acid into the medium plant hormone, heating to 80-100 ℃, stirring at constant temperature for 1-2h, then adding calcium chloride solution, stirring and mixing uniformly to obtain slow-release particle suspension;
(4) Preparing a regeneration medium of the chives: adding 0.1g of 6-benzylaminoadenine, 2g of slow release granule suspension, 10g of sucrose and 10g of agar into 500g of MS culture medium, stirring and heating until the agar is completely dissolved, and cooling to room temperature to obtain the regeneration culture medium of the chives.
Further, in the step (1), the mass ratio of the hydrophobic chitosan, the absolute ethyl alcohol, the dodecylaminopropionic acid and the amphiphilic alginic acid is as follows: (2-3): (8-12): (1-2): (2-3).
Still further, the preparation method of the hydrophobic chitosan in the step (1) is as follows:
dissolving chitosan and 1-hydroxybenzotriazole in deionized water, adding N-acetyl cysteine to be fully dissolved, dropwise adding carbodiimide, adding sodium hydroxide solution, stirring and reacting for 1.5h, and dialyzing for 16-20h to obtain the hydrophobic chitosan.
Still further, the preparation method of the amphiphilic alginic acid in the step (1) comprises the following steps:
adding alginic acid, cholesterol, dicyclohexylcarbodiimide and 2, 2-dimethylolpropionic acid into deionized water, mixing uniformly, and stirring for 2 hours to obtain the amphiphilic alginic acid.
Further, in the step (2), the mass ratio of the naphthalene acetic acid to the indole acetic acid to the deionized water is as follows: (1-2): (1-2): (10-20).
Further, the volume ratio of the amphiphilic chitosan-alginic acid, the plant hormone solution and the calcium chloride solution in the step (3) is as follows: (2-3): (1-2): (0.2-0.3).
The plant hormone in the common plant culture medium is quickly absorbed by plants and cannot be slowly released for a long time, so that the effect is difficult to control, and the rooting rate and the survival rate of rooting culture in the culture process cannot be improved at the same time. The plant hormone is used as a hydrophilic substance, and needs to be wrapped by a hydrophobic or amphiphilic substance so as to achieve the effect of slow release, and the plant tissue culture has high requirements on materials, so that the plant-friendly material is needed to be used for preparing a culture medium. Both chitosan and alginic acid are extracted from plants and used as common plant friendly materials for preparing culture media. Although the hydrophilic phytohormones can be uniformly distributed in the gel, the prepared culture medium has the same problems as the common culture medium in that various phytohormones are rapidly absorbed at the same time. Therefore, the invention prepares the chitosan into the hydrophobic chitosan, prepares the sodium alginate into the amphiphilic alginic acid, and alternately arranges the amphiphilic chitosan and the amphiphilic alginic acid in absolute ethyl alcohol under the action of dodecylaminopropionic acid to form the amphiphilic chitosan-alginic acid. Mixing the amphipathic chitosan-alginic acid with a plant hormone solution under the heating condition and the stirring shearing force, wrapping the plant hormone in the amphipathic chitosan-alginic acid, and then adding a calcium chloride solution to replace alginic acid in the structure with calcium alginate to form a wrapping structure which is more stable to obtain a slow-release particle suspension. The regeneration culture medium of the chives, which is prepared by using the slow-release particle suspension, is slow in release and long in acting time because the plant hormone is wrapped in the amphiphilic chitosan-alginic acid during rooting culture, so that the rooting rate and the survival rate of the chives after rooting culture are improved simultaneously.
The beneficial effects are that:
1. the regeneration culture medium of the chives can effectively control the slow release of plant hormones of naphthalene acetic acid and indole acetic acid, so that the cultivated chives have high rooting rate and high survival rate.
2. The amphipathy chitosan-alginic acid is used for wrapping the naphthylacetic acid and the indoleacetic acid, and the slow-release particles prepared under the action of the calcium chloride solution have good stability, so that the slow release of hormone can be effectively controlled in the regeneration culture process.
Drawings
Fig. 1: inducing and culturing the leek stem node;
fig. 2: a diagram of the induction result of adventitious buds of the chives;
fig. 3: and (5) a root induction result graph of the chives.
Detailed Description
The invention will be described in detail below with reference to examples and figures:
the invention needs to prepare hydrophobic chitosan and amphiphilic alginic acid before preparing a regeneration culture medium of the chives, and specifically comprises the following steps:
the hydrophobic chitosan used in the invention is prepared by the following steps:
after 1g of chitosan and 0.01g of 1-hydroxybenzotriazole are dissolved in 20mL of deionized water, 0.05g of N-acetyl cysteine is added, 5mL of carbodiimide is added dropwise after the mixture is fully dissolved, then 5mL of sodium hydroxide solution is added, stirring reaction is carried out for 1.5h, and the mixture is dialyzed in clear water for 18h to obtain the hydrophobic chitosan.
The amphipathic alginic acid used in the invention is prepared by the following steps:
1g of alginic acid, 0.3g of cholesterol, 0.02g of dicyclohexylcarbodiimide and 0.1g of 2, 2-dimethylolpropionic acid were added to 50mL of deionized water, and the mixture was stirred for 2 hours to obtain amphiphilic alginic acid.
Example 1: preparation of regeneration medium of chives
(1) Preparation of amphiphilic chitosan-alginic acid: weighing 2.5kg of hydrophobic chitosan, adding into 10kg of absolute ethyl alcohol for full dissolution, adding 1.5kg of dodecylaminopropionic acid, continuously stirring for 25min, and then adding 2.5kg of amphiphilic alginic acid, continuously stirring for 1.5h to obtain amphiphilic chitosan-alginic acid for later use;
(2) Preparing a plant hormone solution: 1.5kg of naphthalene acetic acid and 1.5kg of indole acetic acid are respectively weighed and fully dissolved in 15kg of deionized water to obtain a plant hormone solution for standby;
(3) Preparing a slow release particle suspension: weighing 25mL of amphiphilic chitosan-alginic acid, pouring 15mL of plant hormone, heating to 90 ℃, stirring at constant temperature for 1.5h, then adding 2.5mL of calcium chloride solution, stirring and mixing uniformly to obtain slow-release particle suspension;
(4) Preparing a regeneration medium of the chives: adding 0.1g of 6-benzylaminoadenine, 2g of slow release granule suspension, 10g of sucrose and 10g of agar into 500g of MS culture medium, stirring and heating until the agar is completely dissolved, and cooling to room temperature to obtain the regeneration culture medium of the chives.
Example 2: preparation of regeneration medium II of Chinese chives
(1) Preparation of amphiphilic chitosan-alginic acid: weighing 2kg of hydrophobic chitosan, adding into 8kg of absolute ethyl alcohol for full dissolution, adding 2kg of dodecylaminopropionic acid, continuously stirring for 25min, then adding 3kg of amphiphilic alginic acid, and continuously stirring for 1.5h to obtain amphiphilic chitosan-alginic acid for later use;
(2) Preparing a plant hormone solution: 2kg of naphthalene acetic acid and 2kg of indole acetic acid are respectively weighed and fully dissolved in 20kg of deionized water to obtain a plant hormone solution for standby;
(3) Preparing a slow release particle suspension: weighing 20mL of amphiphilic chitosan-alginic acid, pouring 10mL of plant hormone into the mixture, heating to 90 ℃, stirring at constant temperature for 1.5h, then adding 2mL of calcium chloride solution, stirring and mixing uniformly to obtain slow-release particle suspension;
(4) Preparing a regeneration medium of the chives: adding 0.1g of 6-benzylaminoadenine, 2g of slow release granule suspension, 10g of sucrose and 10g of agar into 500g of MS culture medium, stirring and heating until the agar is completely dissolved, and cooling to room temperature to obtain the regeneration culture medium of the chives.
Example 3: preparation of regeneration medium for chives
(1) Preparation of amphiphilic chitosan-alginic acid: weighing 3kg of hydrophobic chitosan, adding into 12kg of absolute ethyl alcohol for full dissolution, adding 1kg of dodecylaminopropionic acid, continuously stirring for 25min, then adding 2kg of amphiphilic alginic acid, and continuously stirring for 1.5h to obtain amphiphilic chitosan-alginic acid for later use;
(2) Preparing a plant hormone solution: 1kg of naphthalene acetic acid and 1kg of indole acetic acid are respectively weighed and fully dissolved in 100kg of deionized water to obtain a plant hormone solution for standby;
(3) Preparing a slow release particle suspension: weighing 30mL of amphiphilic chitosan-alginic acid, pouring 20mL of plant hormone into the mixture, heating to 90 ℃, stirring at constant temperature for 1.5h, then adding 3mL of calcium chloride solution, stirring and mixing uniformly to obtain slow-release particle suspension;
(4) Preparing a regeneration medium of the chives: adding 0.1g of 6-benzylaminoadenine, 2g of slow release granule suspension, 10g of sucrose and 10g of agar into 500g of MS culture medium, stirring and heating until the agar is completely dissolved, and cooling to room temperature to obtain the regeneration culture medium of the chives.
Comparative example 1:
this comparative example is to be compared with example 1, and differs only in that no hydrophobic chitosan is added in step (1), and the rest steps are the same, and the step (1) is specifically as follows:
(1) Preparation of amphiphilic chitosan-alginic acid: adding 2kg of dodecylaminopropionic acid into 10kg of absolute ethyl alcohol, continuously stirring for 25min, then adding 3kg of amphiphilic alginic acid, and continuously stirring for 1.5h to obtain amphiphilic chitosan-alginic acid for later use;
comparative example 2:
this comparative example is to be compared with example 1, and differs only in that dodecylaminopropionic acid is not added in step (1), and the rest steps are the same, and step (1) is specifically as follows:
(1) Preparation of amphiphilic chitosan-alginic acid: weighing 2kg of hydrophobic chitosan, adding the hydrophobic chitosan into 10kg of absolute ethyl alcohol, fully dissolving, stirring for 25min, then adding 3kg of amphiphilic alginic acid, and continuously stirring for 1.5h to obtain amphiphilic chitosan-alginic acid for later use;
comparative example 3:
this comparative example is in contrast to example 1, which differs only in that amphiphilic alginic acid is not added in step (1), and the remaining steps are the same, and step (1) is specifically as follows:
(1) Preparation of amphiphilic chitosan-alginic acid: weighing 2.5kg of hydrophobic chitosan, adding into 10kg of absolute ethyl alcohol for full dissolution, adding 1.5kg of dodecylaminopropionic acid, and continuously stirring for 1.5h to obtain amphiphilic chitosan-alginic acid for later use;
comparative example 4:
this comparative example is to be compared with example 1, and differs only in that no calcium chloride solution is added in step (3), and the rest of the steps are the same, and step (3) is specifically as follows:
(3) Preparing a slow release particle suspension: weighing 25mL of amphiphilic chitosan-alginic acid, pouring 15mL of plant hormone into the mixture, heating to 90 ℃, and stirring at constant temperature for 1.5h to obtain slow-release particle suspension;
comparative example 5:
this comparative example is to be compared with example 1, and differs only in that the hydrophobic chitosan in step (1) is replaced by ordinary chitosan, and the rest steps are the same, and the step (1) is specifically as follows:
(1) Preparation of amphiphilic chitosan-alginic acid: weighing 2.5kg of common chitosan, adding into 10kg of absolute ethyl alcohol for full dissolution, adding 1.5kg of dodecylaminopropionic acid, continuously stirring for 25min, and then adding 2.5kg of amphiphilic alginic acid, continuously stirring for 1.5h to obtain amphiphilic chitosan-alginic acid for later use.
Comparative example 6:
(1) Preparation of amphiphilic chitosan-alginic acid: weighing 2.5kg of hydrophobic chitosan, adding into 10kg of absolute ethyl alcohol for full dissolution, adding 1.5kg of dodecylaminopropionic acid, continuously stirring for 25min, then adding 2.5kg of common calcium alginate, and continuously stirring for 1.5h to obtain the amphiphilic chitosan-alginic acid for later use.
Experiment 1 of regeneration culture of chives
Step one, the explant is disinfected and sterile seedling is obtained
The leek bulbs and leaves are used as explants, outer-layer scales and leaves are removed, the leek bulbs and leaves are rinsed by a detergent for 10min, and the leek bulbs and leaves are rinsed by running water for 2.5h. Transferring to an ultra-clean workbench, sterilizing with 75% ethanol for 30s, washing with sterile water for 2 times, sterilizing with 0.2% mercuric chloride for 0-15min, and washing with sterile water for 5 times.
Step two, callus induction
Preparation of an induction medium: MS 2.22g, NAA 0.1mg, 6-BA 0.1mg, sucrose 15g and agar 15g are respectively weighed, added into deionized water, heated and dissolved, and cooled to room temperature to obtain an induction culture medium.
Inoculating bulbs and leaves in the first step on an induction culture medium, and culturing in dark under the environment condition of 25 ℃ for 16 hours under the illumination time of 3000lux per day. The results of the callus obtained after 40d of culture are shown in FIG. 1.
Step three, callus proliferation
Preparation of proliferation and differentiation medium: MS 4.44g, NAA 0.1mg, 6-BA 1.0mg, sucrose 30g and agar 30g are respectively weighed, added into deionized water, heated and melted, and cooled to room temperature to obtain an induction culture medium.
Dividing the callus of the chives in the second step into 1cm multiplied by 1cm, transferring into proliferation and differentiation culture medium, and illuminating for 16h each day under the environmental condition of 25 ℃ with illumination intensity of 3000lux. The result of the proliferation and differentiation culture for 2 months to obtain adventitious buds of 3cm or more is shown in FIG. 2.
Step four, rooting culture
Preparation of a common rooting medium (i.e., a blank control): MS 2.22g, NAA 0.2mg, IAA 0.1mg, sucrose 15g and agar 8g are respectively weighed and added into deionized water for heating and melting, and then cooled to room temperature to obtain a common rooting culture medium.
The adventitious buds grown to 3cm or more in the third step were transferred to the leek regeneration medium and the general rooting medium (blank control) prepared in example 1 and comparative examples 1 to 6, and cultured for 30d under the environmental conditions of 25 ℃ and illumination time of 16 hours per day and illumination intensity of 3000lux, rooting was induced and the rooting rate and survival rate were recorded, and the results are shown in table 1, wherein the leek sterile regeneration seedlings were obtained by rooting culture using the leek regeneration medium prepared in example 1, and the results are shown in fig. 3.
TABLE 1
| Rooting percentage (%) | Survival rate (%) | |
| Blank control | 42％ | 77％ |
| Example 1 | 97％ | 98％ |
| Comparative example 1 | 47％ | 36％ |
| Comparative example 2 | 29％ | 63％ |
| Comparative example 3 | 31％ | 73％ |
| Comparative example 4 | 79％ | 31％ |
| Comparative example 5 | 78％ | 30％ |
| Comparative example 6 | 50％ | 62％ |
Analysis of results:
as can be seen from comparing the results of example 1 and the blank control in Table 1, the rooting rate of the regenerated culture medium of the leek prepared by the invention for inducing rooting can reach 97%, the survival rate can reach 98%, and the rooting rate is increased by 52% and 21% respectively compared with the blank control. The preparation method is mainly characterized in that the prepared regeneration culture medium of the allium tuberosum has good slow release effect, and can effectively control the release of plant hormones such as naphthalene acetic acid, indole acetic acid, 6-benzylaminoadenine and the like, so that the rooting rate of the allium tuberosum is effectively improved, and the survival rate is also improved.
The data of comparative examples 1 and 1-3 and comparative examples 5-6 show that comparative examples 1-3 are prepared without adding hydrophobic chitosan, dodecylaminopropionic acid and amphiphilic alginic acid, respectively, during step (1); comparative examples 5 to 6 are respectively to change the hydrophobic chitosan used in the process of step (1) into common chitosan and the amphiphilic alginic acid into common sodium alginate. The rooting rate and the survival rate of the comparative examples 1-3 and 5-6 are reduced compared with those of the example 1 after rooting cultivation, mainly because the slow release effect of the leek regeneration medium is that hydrophobic chitosan, dodecylaminopropionic acid and amphiphilic alginic acid are mutually cooperated in the step (1) to obtain amphiphilic chitosan-alginic acid, and the amphiphilic chitosan-alginic acid wraps plant hormones such as naphthalene acetic acid, indole acetic acid and 6-benzylaminoadenine through the special amphiphilic characteristics, so that the amphiphilic chitosan-alginic acid can be slowly released during rooting cultivation.
The data of comparative examples 1 and 4 show that comparative example 4 does not add calcium chloride solution during step (3), and the rooting rate and survival rate of comparative example 4 are both reduced compared with example 1 after rooting cultivation, mainly because the coating structure is unstable after NAA and IAA are coated in amphipathic chitosan-alginic acid, and alginic acid can be converted into calcium alginate after adding calcium chloride solution, thereby improving structural stability, and enabling slow release of plant hormones such as naphthalene acetic acid, indole acetic acid and 6-benzylaminoadenine for a long time during rooting cultivation.
The above embodiments are only for illustrating the technical solution of the present invention and not for limiting the same, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications and equivalents may be made thereto without departing from the spirit and scope of the technical solution of the present invention, which is intended to be covered by the scope of the claims of the present invention. The technology, shape, and construction parts of the present invention, which are not described in detail, are known in the art.
Claims (4)
1. A regeneration medium for chives, which is characterized by comprising the following components: sustained release particle suspension, agar, sucrose, MS medium, 6-benzylaminoadenine;
the preparation method of the chives regeneration medium comprises the following specific steps:
(1) Preparation of amphiphilic chitosan-alginic acid: adding hydrophobic chitosan into absolute ethyl alcohol for full dissolution, adding dodecylaminopropionic acid and amphiphilic alginic acid, and uniformly stirring and mixing to obtain amphiphilic chitosan-alginic acid for later use;
(2) Preparing a plant hormone solution: fully dissolving naphthalene acetic acid and indole acetic acid in deionized water to obtain a plant hormone solution for later use;
(3) Preparing a slow release particle suspension: pouring amphiphilic chitosan-alginic acid into a plant hormone solution, heating and stirring, then adding a calcium chloride solution, stirring and uniformly mixing to obtain a slow-release particle suspension;
(4) Preparing a regeneration medium of the chives: adding 0.1g of 6-benzylaminoadenine, 2g of slow release granule suspension, 10g of sucrose and 10g of agar into 500g of MS culture medium, heating while stirring until the agar is completely dissolved, and cooling to room temperature to obtain a regeneration culture medium of the chives;
the preparation method of the hydrophobic chitosan in the step (1) comprises the following steps:
dissolving chitosan and 1-hydroxybenzotriazole in deionized water, adding N-acetylcysteine to be fully dissolved, dropwise adding carbodiimide, then adding sodium hydroxide solution, stirring and reacting for 1.5 hours, and dialyzing to obtain hydrophobic chitosan;
the preparation method of the amphipathic alginic acid in the step (1) comprises the following steps:
adding alginic acid, cholesterol, dicyclohexylcarbodiimide and 2, 2-dimethylolpropionic acid into deionized water, mixing uniformly, and stirring for 2 hours to obtain the amphiphilic alginic acid.
2. The regeneration medium for chives according to claim 1, wherein the mass ratio of the hydrophobic chitosan, the absolute ethyl alcohol, the dodecylaminopropionic acid and the amphiphilic alginic acid in the step (1) is (2-3): (8-12): (1-2): (2-3).
3. The regeneration medium for chives according to claim 2, wherein in the step (2), the mass ratio of the naphthalene acetic acid, the indole acetic acid and the deionized water is (1-2): (1-2): (10-20).
4. The regeneration medium for chives according to claim 3, wherein the volume ratio of the amphiphilic chitosan-alginic acid, the plant hormone solution and the calcium chloride solution in the step (3) is (2-3): (1-2): (0.2-0.3).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310017469.5A CN115812604B (en) | 2023-01-06 | 2023-01-06 | Leek regeneration medium and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310017469.5A CN115812604B (en) | 2023-01-06 | 2023-01-06 | Leek regeneration medium and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115812604A CN115812604A (en) | 2023-03-21 |
| CN115812604B true CN115812604B (en) | 2023-08-29 |
Family
ID=85520286
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310017469.5A Active CN115812604B (en) | 2023-01-06 | 2023-01-06 | Leek regeneration medium and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115812604B (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004049164A (en) * | 2002-07-23 | 2004-02-19 | Japan Atom Energy Res Inst | Method for tissue culture of plant using irradiated polysaccharide |
| WO2005012507A1 (en) * | 2003-07-25 | 2005-02-10 | The University Of Melbourne | Production of plant secondary metabolites using adsorption and elicitation in cell suspension culture |
| CN114303460B (en) * | 2007-08-31 | 2009-07-01 | 陕西佰美基因股份有限公司 | Method for preparing ring-opening saccharide cross-linked hemoglobin for red blood cell substitute |
| CN107047311A (en) * | 2017-05-11 | 2017-08-18 | 平顶山市平丰种业有限责任公司 | A kind of leek Unfertilized Ovaries in-vitro culture method |
| CN108084139A (en) * | 2017-12-15 | 2018-05-29 | 柳州旭至自动化科技有限公司 | A kind of onion anthocyanidin preparation method |
| CN109554360A (en) * | 2019-01-25 | 2019-04-02 | 上海应用技术大学 | A method of thallus is embedded using sodium alginate composite material |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015126944A1 (en) * | 2014-02-18 | 2015-08-27 | Henkin Robert I | Methods and compositions for diagnosing and treating loss and/or distortion of taste or smell |
-
2023
- 2023-01-06 CN CN202310017469.5A patent/CN115812604B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004049164A (en) * | 2002-07-23 | 2004-02-19 | Japan Atom Energy Res Inst | Method for tissue culture of plant using irradiated polysaccharide |
| WO2005012507A1 (en) * | 2003-07-25 | 2005-02-10 | The University Of Melbourne | Production of plant secondary metabolites using adsorption and elicitation in cell suspension culture |
| CN114303460B (en) * | 2007-08-31 | 2009-07-01 | 陕西佰美基因股份有限公司 | Method for preparing ring-opening saccharide cross-linked hemoglobin for red blood cell substitute |
| CN107047311A (en) * | 2017-05-11 | 2017-08-18 | 平顶山市平丰种业有限责任公司 | A kind of leek Unfertilized Ovaries in-vitro culture method |
| CN108084139A (en) * | 2017-12-15 | 2018-05-29 | 柳州旭至自动化科技有限公司 | A kind of onion anthocyanidin preparation method |
| CN109554360A (en) * | 2019-01-25 | 2019-04-02 | 上海应用技术大学 | A method of thallus is embedded using sodium alginate composite material |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115812604A (en) | 2023-03-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105053559A (en) | Active dry yeast for accelerating growth and development of animals and preparation method of active dry yeast | |
| CN108040875A (en) | A kind of screening collocation method of the culture medium of efficient induction embryo callus | |
| CN107988089B (en) | Preparation method of saccharomyces boulardii preparation and saccharomyces boulardii preparation | |
| CN111034616B (en) | Broccoli hairy root high sulforaphane glycoside and sulforaphane synthesis release culture method | |
| CN107970446B (en) | Piglet diarrhea-resistant high-immunity whole egg powder microcapsule and preparation method and application thereof | |
| CN115812604B (en) | Leek regeneration medium and preparation method thereof | |
| CN103039360B (en) | Method for quickly propagating leeka through tissue culture | |
| CN107148911A (en) | A kind of sweet potato stem tip detoxification solid medium and preparation method thereof | |
| CN106995347A (en) | A kind of Flower Plants in Hydroponic Culture nutrient solution and preparation method thereof | |
| CN106718042A (en) | A kind of method of utilization bamboo rat excrement Xinbao mushroom culturing | |
| KR100505936B1 (en) | Sugar cane production | |
| CN112876305A (en) | Microbial compound fertilizer and preparation method thereof | |
| Kim et al. | High frequency plant regeneration of garlic (Allium sativum L.) calli immobilized in calcium alginate gel | |
| CN106665364A (en) | Lilium brownii var.viridulum filial generation bulb expanding, developing and rooting method | |
| CN113880638A (en) | Rice seed soaking agent based on kitchen biogas slurry and preparation method and application thereof | |
| CN103004596B (en) | Method for quickly obtaining garlic seedlings by using intermittent immersion cultivation mode | |
| CN106258985A (en) | A kind of culture medium extending tissue culture seedlings of bananas storage life and cultural method | |
| CN107853178B (en) | Method for inducing embryogenic callus | |
| CN111194696A (en) | In-vitro plant material tissue culture medium with bacteria and tissue culture method | |
| CN110679765A (en) | Feed additive for relieving piglet stress | |
| CN1260184C (en) | Preparation method of high efficient biological organic fertilizer | |
| CN112715774B (en) | Young poultry feed and preparation method thereof | |
| CN108040881A (en) | A kind of dragon fruit synchronization tissue culture culture medium and the cultural method of plant | |
| CN104388379A (en) | Method for obtaining sugarcane calluses from spermine | |
| CN115812595B (en) | Preparation method of artificial seeds of fritillaria cirrhosa |
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 |