CN111837952B - Peucedanum praeruptorum cluster bud culture medium with bud tip as explant - Google Patents
Peucedanum praeruptorum cluster bud culture medium with bud tip as explant Download PDFInfo
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- CN111837952B CN111837952B CN202010668558.2A CN202010668558A CN111837952B CN 111837952 B CN111837952 B CN 111837952B CN 202010668558 A CN202010668558 A CN 202010668558A CN 111837952 B CN111837952 B CN 111837952B
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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 discloses a peucedanum cluster bud culture medium with bud tips as explants, wherein each 1000mL of the culture medium contains 1.5-2.5mg of 6-benzylamino adenine, 0.5-1.5mg of kinetin, 0.2-0.4mg of indoleacetic acid, 160mg of coconut milk, 3-5g of agar powder, 28-34g of cane sugar and the balance of a basic culture medium. The peucedanum praeruptorum clustered bud culture medium taking the bud tips as the explants can greatly improve the induction rate of the peucedanum praeruptorum clustered bud induction taking the bud tips as the explants, improve the production efficiency and save the production cost.
Description
Technical Field
The invention relates to the technical field of peucedanum praeruptorum tissue culture, in particular to a peucedanum praeruptorum cluster bud culture medium taking bud tips as explants.
Background
Radix Peucedani is a perennial herb of Umbelliferae, and has irregular cylindrical, conical or spindle-shaped root, and dark brown or gray yellow surface. The peucedanum root is a common Chinese medicinal material, is taken as a medicine by dry roots, has the effects of depressing qi, reducing phlegm, dispelling wind and clearing heat, and is mainly used for treating phlegm-heat, dyspnea with fullness, yellow and thick sputum, wind-heat cough with excessive phlegm and other symptoms.
At present, the artificial propagation of the peucedanum praeruptorum is mainly carried out by adopting a sowing and tissue culture mode, and the prior tissue culture technology of the peucedanum praeruptorum mostly takes leaves, petioles and stem sections as explants, so that the induction and differentiation efficiency is low, and the peucedanum praeruptorum is easy to be infected with viruses.
Disclosure of Invention
Based on the technical problems in the background art, the invention provides a peucedanum clustered bud culture medium taking bud tips as explants.
Every 1000mL of the culture medium of the cluster buds of the peucedanum praeruptorum which takes bud tips as explants contains 1.5-2.5mg of 6-benzylamino adenine, 0.5-1.5mg of kinetin, 0.2-0.4mg of indoleacetic acid, 160mg of coconut milk, 3-5g of agar powder, 28-34g of cane sugar and the balance of basal culture medium; the coconut milk is collected from unripe coconut fruit.
Preferably, each 1000mL of the culture medium contains 1.8-2.2mg of 6-benzylamino adenine, 0.8-1.2mg of kinetin, 0.25-0.35mg of indoleacetic acid, 150mg of coconut milk, 3.5-4.5g of agar powder, 30-32g of cane sugar and the balance of basal medium.
Preferably, each 1000mL of the culture medium contains 2.0mg of 6-benzylamino adenine, 1.0mg of kinetin, 0.3mg of indoleacetic acid, 140mg of coconut milk, 4.0g of agar powder, 31g of cane sugar and the balance of a basal culture medium.
Preferably, the coconut milk is collected from immature coconut fruit grown for 5-7 months.
The basic culture medium can be a conventional basic culture medium for plant tissue culture, and preferably, the basic culture medium is an MS culture medium.
The invention has the following beneficial effects:
the peucedanum cluster bud culture medium with bud tips as explants contains active ingredients such as 6-benzylamino adenine, kinetin, indoleacetic acid, coconut milk and the like. Wherein, the indoleacetic acid can promote cell growth; kinetin and 6-benzylamino adenine can promote cell division and differentiation; the coconut milk is liquid endosperm of coconut, contains various nutrient substances and regulating substances required by plant growth, has multiple functions of promoting cell growth, division and differentiation, and particularly has remarkable promoting effect on the coconut milk collected from immature coconut fruits growing for 5-7 months, and the mature coconut fruits consume part of the nutrient substances in the coconut milk, so that the plant growth regulator for promoting cell growth and division has low content and poor promoting effect; the components can play a synergistic role, and better promote the bud tips of the peucedanum praeruptorum to induce cluster buds with calluses, thereby obviously improving the induction rate of the cluster buds of the peucedanum praeruptorum. The invention provides the peucedanum cluster bud culture medium taking bud tips as explants, which has the advantages of high induction rate, simple material selection, low virus infection rate, improved production efficiency and production cost saving.
Detailed Description
The technical solution of the present invention will be described in detail below with reference to specific examples.
In the following examples and comparative examples, coconut milk is the liquid endosperm of immature coconut and is collected from coconut fruit grown for 6 months.
Example 1
Preparing a peucedanum cluster bud culture medium, which comprises the following components: each 1000mL of the culture medium contains 1.5mg of 6-benzylamino adenine, 0.5mg of kinetin, 0.2mg of indoleacetic acid, 120mg of coconut milk, 3g of agar powder, 28g of cane sugar and the balance of MS culture medium.
Example 2
Preparing a peucedanum cluster bud culture medium, which comprises the following components: each 1000mL of the culture medium contains 2.5mg of 6-benzylamino adenine, 1.5mg of kinetin, 0.4mg of indoleacetic acid, 160mg of coconut milk, 5g of agar powder, 34g of cane sugar and the balance of MS culture medium.
Example 3
Preparing a peucedanum cluster bud culture medium, which comprises the following components: each 1000mL of the culture medium contains 1.8mg of 6-benzylamino adenine, 0.8mg of kinetin, 0.35mg of indoleacetic acid, 130mg of coconut milk, 4g of agar powder, 31g of cane sugar and the balance of MS culture medium.
Example 4
Preparing a peucedanum cluster bud culture medium, which comprises the following components: each 1000mL of the culture medium contains 1.8mg of 6-benzylamino adenine, 1.0mg of kinetin, 0.25mg of indoleacetic acid, 140mg of coconut milk, 4g of agar powder, 31g of cane sugar and the balance of MS culture medium.
Example 5
Preparing a peucedanum cluster bud culture medium, which comprises the following components: each 1000mL of the culture medium contains 1.8mg of 6-benzylamino adenine, 1.2mg of kinetin, 0.3mg of indoleacetic acid, 150mg of coconut milk, 4g of agar powder, 31g of cane sugar and the balance of MS culture medium.
Example 6
Preparing a peucedanum cluster bud culture medium, which comprises the following components: each 1000mL of the culture medium contains 2.0mg of 6-benzylamino adenine, 0.8mg of kinetin, 0.25mg of indoleacetic acid, 130mg of coconut milk, 4g of agar powder, 31g of cane sugar and the balance of MS culture medium.
Example 7
Preparing a peucedanum cluster bud culture medium, which comprises the following components: each 1000mL of the culture medium contains 2.0mg of 6-benzylamino adenine, 1.0mg of kinetin, 0.3mg of indoleacetic acid, 140mg of coconut milk, 4.0g of agar powder, 31g of cane sugar and the balance of MS culture medium.
Example 8
Preparing a peucedanum cluster bud culture medium, which comprises the following components: each 1000mL of the culture medium contains 2.0mg of 6-benzylamino adenine, 1.0mg of kinetin, 0.35mg of indoleacetic acid, 150mg of coconut milk, 4.0g of agar powder, 31g of cane sugar and the balance of MS culture medium.
Example 9
Preparing a peucedanum cluster bud culture medium, which comprises the following components: each 1000mL of the culture medium contains 2.2mg of 6-benzylamino adenine, 0.8mg of kinetin, 0.3mg of indoleacetic acid, 130mg of coconut milk, 4.0g of agar powder, 31g of cane sugar and the balance of MS culture medium.
Example 10
Preparing a peucedanum cluster bud culture medium, which comprises the following components: each 1000mL of the culture medium contains 2.2mg of 6-benzylamino adenine, 1.0mg of kinetin, 0.35mg of indoleacetic acid, 140mg of coconut milk, 4.0g of agar powder, 31g of cane sugar and the balance of MS culture medium.
Example 11
Preparing a peucedanum cluster bud culture medium, which comprises the following components: each 1000mL of the culture medium contains 2.2mg of 6-benzylamino adenine, 1.2mg of kinetin, 0.25mg of indoleacetic acid, 150mg of coconut milk, 4.0g of agar powder, 31g of cane sugar and the balance of MS culture medium.
Comparative example 1
Preparing a peucedanum cluster bud culture medium, which comprises the following components: each 1000mL of the culture medium contains 2.0mg of 6-benzylamino adenine, 1.0mg of kinetin, 0.3mg of indoleacetic acid, 4.0g of agar powder, 31g of cane sugar and the balance of MS culture medium.
Comparative example 2
Preparing a peucedanum cluster bud culture medium, which comprises the following components: each 1000mL of the culture medium contains 2.0mg of 6-benzylamino adenine, 1.0mg of kinetin, 140mg of coconut milk, 4.0g of agar powder, 31g of cane sugar and the balance of MS culture medium.
Comparative example 3
Preparing a peucedanum cluster bud culture medium, which comprises the following components: each 1000mL of the culture medium contains 2.0mg of 6-benzylamino adenine, 0.3mg of indoleacetic acid, 140mg of coconut milk, 4.0g of agar powder, 31g of cane sugar and the balance of MS culture medium.
Comparative example 4
Preparing a peucedanum cluster bud culture medium, which comprises the following components: each 1000mL of the culture medium contains 1.0mg of kinetin, 0.3mg of indoleacetic acid, 140mg of coconut milk, 4.0g of agar powder, 31g of cane sugar and the balance of MS culture medium.
The medium composition of examples 1-11 and comparative examples 1-4 is shown in Table 1:
TABLE 1 culture Medium Components of examples 1-11 and comparative examples 1-4
Test examples
The induction of multiple shoots was carried out using the medium of examples 1 to 11 and comparative examples 1 to 4, with the shoot tips as explants. The method comprises the following specific steps: washing strong terminal bud of radix Peucedani with running water for 0.5h, soaking in 75% ethanol for 5s under aseptic condition, soaking in 0.1% mercuric chloride solution for 10-15min, washing with sterile water for 3-5 times, stripping 0.8-1.0mm bud tip, inoculating on culture medium, and culturing under 24-26 deg.C, illumination intensity of 1200lx, and illumination time of 12h per day. Subculture for 1 time for 30 days, and recording the induction condition of cluster buds of the sterilized test-tube plantlet of radix peucedani when culturing for 60 days. The induction rate calculation method comprises the following steps: the induction rate is the number of bud tips inducing the cluster buds/the total number of inoculated bud tips.
The induction test results are shown in table 2:
TABLE 2 induction rates of the media of examples 1 to 11 and comparative examples 1 to 4
From the test results in table 2, it can be seen that the peucedanum cluster bud culture medium using bud tips as explants according to the present invention can successfully induce cluster buds using bud tips as explants, and the induction rate can reach 96.8%, wherein the induction rate of peucedanum cluster buds using bud tips as explants can be greatly improved by adding coconut milk in the culture medium.
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 (3)
1. A peucedanum cluster bud culture medium taking bud tips as explants is characterized in that every 1000mL of the culture medium contains 2.0mg of 6-benzylamino adenine, 1.0mg of kinetin, 0.3mg of indoleacetic acid, 140mg of coconut milk, 4.0g of agar powder, 31g of cane sugar and the balance of basal culture medium; and (4) stripping bud tips, inoculating the bud tips on a culture medium, and performing cluster bud induction.
2. The culture medium for the clustered shoots of peucedanum praeruptorum with shoot tips as explants according to claim 1, wherein the coconut milk is collected from immature coconut fruits growing for 5-7 months.
3. The culture medium for the clustered shoots of peucedanum praeruptorum with shoot tips as explants according to claim 1, wherein the basic culture medium is MS culture medium.
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| CN115380830B (en) * | 2022-10-08 | 2023-08-01 | 浙江大学 | Propagation method of adventitious buds of peucedanum praeruptorum dunn |
| CN115777536B (en) * | 2022-11-30 | 2023-08-18 | 中南民族大学 | Method for establishing efficient regeneration system by utilizing stems of peucedanum praeruptorum dunn |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107223564A (en) * | 2017-06-06 | 2017-10-03 | 江苏大学 | A kind of root of purple-flowered peucedanum plant regeneration system method for building up |
| CN108450329A (en) * | 2018-02-09 | 2018-08-28 | 鲁东大学 | A kind of method of diversiform-leaved poplar blade high-frequency induction regeneration plant |
| WO2019006470A1 (en) * | 2017-06-30 | 2019-01-03 | Booshoot Llc | Media for rapid and reliable tissue culturing of plants |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107223564A (en) * | 2017-06-06 | 2017-10-03 | 江苏大学 | A kind of root of purple-flowered peucedanum plant regeneration system method for building up |
| WO2019006470A1 (en) * | 2017-06-30 | 2019-01-03 | Booshoot Llc | Media for rapid and reliable tissue culturing of plants |
| CN108450329A (en) * | 2018-02-09 | 2018-08-28 | 鲁东大学 | A kind of method of diversiform-leaved poplar blade high-frequency induction regeneration plant |
Non-Patent Citations (2)
| Title |
|---|
| Direct somatic embryogenesis and plant regeneration in Peucedanum oreoselinum (L.) Moench .;A. COSTE et al.;《Romanian Biotechnological Letters》;20111231;第16卷(第4期);第6450-6459页 * |
| 正交设计法优选紫花前胡组织培养基;彭菲 等;《中药材》;20010731;第24卷(第7期);第471-472页 * |
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