CN110972946B - Subculture method of rabdosia amethystoides - Google Patents
Subculture method of rabdosia amethystoides Download PDFInfo
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- CN110972946B CN110972946B CN201911316789.0A CN201911316789A CN110972946B CN 110972946 B CN110972946 B CN 110972946B CN 201911316789 A CN201911316789 A CN 201911316789A CN 110972946 B CN110972946 B CN 110972946B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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Abstract
The invention belongs to the technical field of plant tissue culture, and discloses a pulse-developing rabdosia amethystoides subculture method. The subculture method of rabdosia amethystoides comprises the following steps: (1) selecting aseptic seedlings: selecting aseptic seedlings of the rabdosia nervosa with the plant height of 2-3 cm from the starting culture medium; (2) subculturing of aseptic seedlings: transferring the aseptic seedlings to a subculture medium for multiple subcultures. The invention provides a method for subculturing rabdosia amethystoides for the first time, which has the following advantages: the multiplication coefficient is high, and the average multiplication coefficient reaches 8-15; can effectively reduce the phenomenon of serious vitrification after multiple subcultures. The formula not only effectively reduces the production cost, brings higher economic benefit in large-scale production, and provides possibility for industrial production of the pulse-taking rabdosia amethystoides by reasonably adjusting the hormone concentration and the culture medium formula.
Description
Technical Field
The invention belongs to the technical field of plant tissue culture, and discloses a pulse-developing rabdosia amethystoides subculture method.
Technical Field
The Rabdosia amethystoides is also called Hemsl-leaf snake head, Bupleurum falcatum, Rabdosia amethystoides, etc., and belongs to Labiatae, Ocimum basilicum, Rabdosia family species. The whole herbs are used as the medicine, collected in summer and autumn, cleaned and fresh or dried in the sun, slightly pungent, bitter and cold in taste, mainly used for clearing heat, promoting diuresis and detoxifying, and used for acute icteric hepatitis and venomous snake bite; it is used externally to treat burns, scalds, venomous snake bite, impetigo, eczema and skin pruritus.
The traditional asexual propagation of the rabdosia nervosa has low survival rate, low propagation coefficient and slow seedling forming, and the character variation exists in seed propagation, which is not beneficial to keeping the excellent characters of female parent. At present, the requirements of the seedling market are difficult to meet, and a tissue culture technology is adopted, so that an excellent single plant is selected as an explant, the stable inheritance of the hereditary character is ensured, and the seedling quality is improved. In the past, a report related to subculture of rabdosia nervosa is lacked. The phenomena of low proliferation coefficient, serious vitrification and the like exist in the tissue culture process.
In conclusion, the establishment of a tissue culture and rapid propagation method of rabdosia amethystoides is a technical problem which needs to be solved urgently at present.
Disclosure of Invention
The invention aims to solve the technical problem of providing a pulse-developing rabdosia amethystoides subculture method aiming at the defects of the prior art. Through multiple tests, the subculture method of the rabdosia nervosa is summarized, the multiplication coefficient is high and is 8-15, the possibility of providing a large number of high-quality rabdosia nervosa seedlings in the market is provided, and a road is paved for large-scale and industrial production of the rabdosia nervosa seedlings.
In order to solve the technical problems, the invention adopts the technical scheme that: a vein-revealing rabdosia successive transfer culture method is characterized by comprising the following steps of:
a hyponow culture method of rabdosia nervosa comprises selection of aseptic seedlings and subculture of the aseptic seedlings, and comprises the following specific steps:
(1) selecting aseptic seedlings: selecting aseptic seedlings of the rabdosia nervosa which have regular growth vigor and 2-3 cm plant height and are cultured for 5-7 days from a starting culture medium as a subculture material;
(2) subculturing of aseptic seedlings: transferring the aseptic seedlings to a subculture medium for multiple subcultures;
the subculture medium comprises 0.5-1.5 mg/L of MS +6-BA, 28-32 g/L of white granulated sugar and 4-6 g/L of agar, and the pH value is 5.6-6.0.
The whole culture process is carried out in a plant tissue culture room, a fluorescent lamp is adopted for irradiation, the illumination time is 10-14 h/d, the illumination intensity is 2300-2700 lx, and the culture temperature is 23-25 ℃.
And sterilizing the subculture medium by using an autoclave for 18-22 min at 120-122 ℃.
Compared with the prior art, the invention has the following advantages:
the invention provides a method for subculturing rabdosia amethystoides for the first time, the subculturing period is controlled within 25-30 days, the multiplication coefficient is high, and the average multiplication coefficient reaches 8-15; the phenomenon of serious vitrification after multiple subcultures can be effectively reduced, the formula not only effectively reduces the production cost by reasonably adjusting the hormone concentration and the formula of the culture medium, but also brings higher economic benefit in large-scale production and provides possibility for industrial production of the rabdosia amethystoides.
Detailed Description
Example 1
A pulse-developing rabdosia amethystoides subculture method comprises the following specific steps:
(1) selecting aseptic seedlings: selecting aseptic seedlings of the rabdosia nervosa which have regular growth vigor and 2-3 cm plant height and are cultured for 5-7 days from a starting culture medium as a subculture material;
(2) subculturing of aseptic seedlings: transferring the aseptic seedlings to a subculture medium for multiple subcultures, wherein the proliferation medium comprises MS +6-BA 0.5mg/L + white granulated sugar 30g/L + agar 5/L, and the pH value is 5.8. The whole culture process is carried out in a plant tissue culture room, a fluorescent lamp is adopted for irradiation, the illumination time is 12h/d, the illumination intensity is 2500lx, and the culture temperature is 24 ℃. The proliferation medium was sterilized by autoclaving and was sterilized at 121 ℃ for 20 min.
After 25 days of culture, the proliferating mother seedlings without roots, with more lateral branches and higher proliferating proportion are screened out, and the proliferating coefficient is 8.
Example 2
A pulse-developing rabdosia amethystoides subculture method comprises the following specific steps:
(1) selecting aseptic seedlings: selecting aseptic seedlings of the rabdosia nervosa which have regular growth vigor and 2-3 cm plant height and are cultured for 5-7 days from a starting culture medium as a subculture material;
(2) subculturing of aseptic seedlings: transferring the aseptic seedlings to a subculture medium for multiple subcultures, wherein the proliferation medium comprises MS +6-BA 1mg/L + white granulated sugar 28g/L + agar 6/L, and the pH value is 5.6. The whole culture process is carried out in a plant tissue culture room, a fluorescent lamp is adopted for irradiation, the illumination time is 10h/d, the illumination intensity is 2600lx, and the culture temperature is 25 ℃. The proliferation medium was sterilized by autoclaving and was sterilized at 122 ℃ for 21 min.
After culturing for 26 days, the proliferating mother seedlings without roots, with more lateral branches and higher proliferating proportion are screened out, and the proliferating coefficient is 12.
Example 3
A pulse-developing rabdosia amethystoides subculture method comprises the following specific steps:
(1) selecting aseptic seedlings: selecting aseptic seedlings of the rabdosia nervosa which have regular growth vigor and 2-3 cm plant height and are cultured for 5-7 days from a starting culture medium as a subculture material;
(2) subculturing of aseptic seedlings: transferring the aseptic seedlings to a subculture medium for multiple subcultures, wherein the proliferation medium comprises MS +6-BA 1.5mg/L + white granulated sugar 32g/L + agar 6/L, and the pH value is 6. The whole culture process is carried out in a plant tissue culture room, a fluorescent lamp is adopted for irradiation, the illumination time is 14h/d, the illumination intensity is 2700lx, and the culture temperature is 23 ℃. The proliferation medium was sterilized by autoclaving and at 120 ℃ for 18 min.
After 30 days of culture, the proliferating mother seedlings without roots, with more lateral branches and higher proliferating proportion are screened out, and the proliferating coefficient is 15.
Example 4
A pulse-developing rabdosia amethystoides subculture method comprises the following specific steps:
(1) selecting aseptic seedlings: selecting aseptic seedlings of the rabdosia nervosa which have regular growth vigor and 2-3 cm plant height and are cultured for 5-7 days from a starting culture medium as a subculture material;
(2) subculturing of aseptic seedlings: transferring the aseptic seedlings to a subculture medium for multiple subcultures, wherein the proliferation medium comprises MS +6-BA 1.2mg/L + white granulated sugar 29g/L + agar 5/L, and the pH value is 5.8. The whole culture process is carried out in a plant tissue culture room, a fluorescent lamp is adopted for irradiation, the illumination time is 10h/d, the illumination intensity is 2300lx, and the culture temperature is 25 ℃. The proliferation medium was sterilized by autoclaving and was sterilized at 121 ℃ for 22 min.
After 28 days of culture, the proliferating mother seedlings without roots, with more lateral branches and higher proliferating proportion are screened out, and the proliferating coefficient is 14.
Example 5
A pulse-developing rabdosia amethystoides subculture method comprises the following specific steps:
(1) selecting aseptic seedlings: selecting aseptic seedlings of the rabdosia nervosa which have regular growth vigor and 2-3 cm plant height and are cultured for 5-7 days from a starting culture medium as a subculture material;
(2) subculturing of aseptic seedlings: transferring the aseptic seedlings to a subculture medium for multiple subcultures, wherein the proliferation medium comprises MS +6-BA 1.5mg/L + white granulated sugar 30g/L + agar 5/L, and the pH value is 5.8. The whole culture process is carried out in a plant tissue culture room, a fluorescent lamp is adopted for irradiation, the illumination time is 12h/d, the illumination intensity is 2500lx, and the culture temperature is 24 ℃. The proliferation medium was sterilized by autoclaving and was sterilized at 121 ℃ for 20 min.
After 25 days of culture, the proliferating mother seedlings without roots, with more lateral branches and higher proliferating proportion are screened out, and the proliferating coefficient is 13.
The above description is only for the preferred embodiment of the present invention, and is not intended to limit the present invention in any way. Any simple modification, change and equivalent changes of the above embodiments according to the principles of the present invention are still within the protection scope of the technical solution of the present invention.
Claims (2)
1. A hyponow culture method of rabdosia amethystoides is characterized by comprising selection of aseptic seedlings and subculture of the aseptic seedlings, and comprises the following specific steps:
(1) selecting aseptic seedlings: selecting aseptic seedlings of the rabdosia nervosa which have regular growth vigor and 2-3 cm plant height and are cultured for 5-7 days from a starting culture medium as a subculture material;
(2) subculturing of aseptic seedlings: transferring the aseptic seedlings to a subculture medium for multiple subcultures, wherein the whole culture process is carried out in a plant tissue culture room, a fluorescent lamp is adopted for irradiation, the illumination time is 12h/d, the illumination intensity is 2500lx, the culture temperature is 24 ℃, the subculture period is controlled within 25-30 days, and the average multiplication coefficient of the obtained multiplication parent seedlings reaches 8-15;
the subculture medium comprises 0.5-1.5 mg/L of MS +6-BA, 30g/L of white granulated sugar and 5g/L of agar, and the pH value is 5.8.
2. The method for subculturing rabdosia nervosa as claimed in claim 1, wherein the subculturing medium is sterilized by an autoclave at 121 ℃ for 20 min.
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| CN109042119A (en) * | 2018-08-20 | 2018-12-21 | 王远能 | The artificial method for planting of glaucocalyx rabdosia herb |
| CN109362566A (en) * | 2018-11-27 | 2019-02-22 | 钟天路 | A kind of Rabdosia amethystoides tissue culture and rapid propagation method |
| CN110972829A (en) * | 2019-12-19 | 2020-04-10 | 美尚生态景观股份有限公司 | Method for transplanting and hardening tissue culture seedlings of rabdosia nervosa |
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| WO2006012326A1 (en) * | 2004-06-25 | 2006-02-02 | E.I. Dupont De Nemours And Company | Delta-8 desaturase and its use in making polyunsaturated fatty acids |
| CN102242163A (en) * | 2011-04-29 | 2011-11-16 | 郑州大学 | Biological transformation and synthesis method of rubescensin |
| CN109042119A (en) * | 2018-08-20 | 2018-12-21 | 王远能 | The artificial method for planting of glaucocalyx rabdosia herb |
| CN109362566A (en) * | 2018-11-27 | 2019-02-22 | 钟天路 | A kind of Rabdosia amethystoides tissue culture and rapid propagation method |
| CN110972829A (en) * | 2019-12-19 | 2020-04-10 | 美尚生态景观股份有限公司 | Method for transplanting and hardening tissue culture seedlings of rabdosia nervosa |
Non-Patent Citations (3)
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| Adventitious shoot regeneration from leaf explants of the valuable medicinal herb Plectranthus barbatus Andrew;Thangavel等;《AFRICAN JOURNAL OF BIOTECHNOLOGY》;20111230;第10卷(第43期);第8563页 * |
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| 四种植物生长调节剂对线纹香茶菜组织培养的影响;黄群声等;《亚热带植物科学》;20030330;第32卷(第1期);第1.2节、第2.2节 * |
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