CN111869565B - Culture method for expanding propagation of pseudo-ginseng green embryogenic callus - Google Patents
Culture method for expanding propagation of pseudo-ginseng green embryogenic callus Download PDFInfo
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- CN111869565B CN111869565B CN202010687824.6A CN202010687824A CN111869565B CN 111869565 B CN111869565 B CN 111869565B CN 202010687824 A CN202010687824 A CN 202010687824A CN 111869565 B CN111869565 B CN 111869565B
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- 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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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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 culture method for expanding propagation of pseudo-ginseng green embryogenic callus, which relates to the technical field of callus culture and comprises the following steps: step A: preparing pseudo-ginseng callus; and (B) step (B): preparing a liquid culture medium; step C: and (5) culturing a paper bridge. According to the culture method for expanding propagation of pseudo-ginseng green embryogenic callus, the liquid culture medium prepared by the formula is matched with the paper bridge for culturing pseudo-ginseng callus, and the pseudo-ginseng callus can quickly and continuously absorb the culture solution in the liquid culture medium by the culture method, so that the nutrient absorption of the pseudo-ginseng callus is stable, balanced and efficient, the effect of quickly expanding propagation of the pseudo-ginseng callus is achieved, and the pseudo-ginseng callus is enabled to grow more stably, and has high rising speed, healthy callus and more excellent growth situation by the aid of the liquid culture medium composed of four hormones of 2,4-D, 6-BA, NAA and IBA.
Description
Technical Field
The invention relates to the technical field of callus culture, in particular to a culture method for expanding propagation of pseudo-ginseng green embryogenic callus.
Background
Notoginseng radix is a plant of Araliaceae, and is mainly produced in Yunnan mountain state, so it is named Wenshan Notoginseng radix. Sweet, bitter and warm in nature, has the effects of stopping bleeding, relieving pain, promoting blood circulation and removing blood stasis, and has a conical or cylindrical shape of the main root of pseudo-ginseng. The surface is grey brown or grey yellow, and has intermittent longitudinal wrinkles and branch root marks; the top end is provided with a stem mark, and the periphery is provided with a tumor-shaped protrusion; the weight is firm, the section is gray green, yellow green or gray white, and the wood parts are slightly arranged in a radial shape; the ribs are cylindrical or conical; the shearing mouth is in an irregular shrinkage block shape or a strip shape, the surface is provided with a plurality of obvious stem marks and ring patterns, the center of the section is gray or white, and the edges are dark green or gray.
Notoginseng radix is called hemostatic drug, and is suitable for bleeding with blood stasis. Pseudo-ginseng has a strong hemostatic effect, and has obvious hemostatic effect on different animals, different administration routes and different preparations; notoginseng radix has effects of promoting blood circulation, dispelling blood stasis, resisting blood platelet aggregation, and resisting thrombosis; modern researches prove that the pseudo-ginseng has the function of replenishing blood; notoginseng radix injection can remarkably promote recovery of erythrocyte, reticulocyte and hemoglobin, and has no need for market.
Through the callus culture technology, the seedlings of the pseudo-ginseng can be quickly cultivated and propagated, so that the purpose of efficiently cultivating and utilizing the pseudo-ginseng is achieved.
In China, patent application number: CN201810872944.6 discloses a culture system and a culture method for rapid propagation of pseudo-ginseng callus, which are not balanced and stable enough in the supply of culture solution to pseudo-ginseng callus during the actual propagation of pseudo-ginseng callus, have low overall nutrient absorption efficiency of pseudo-ginseng callus, and have long growth cycle, and cannot achieve the effect of rapid propagation of pseudo-ginseng callus.
Therefore, it is necessary to provide a culture method for expanding and propagating pseudo-ginseng green embryogenic callus to solve the above problems.
Disclosure of Invention
The invention aims to provide a culture method for expanding propagation of pseudo-ginseng green embryogenic callus, which aims to solve the problems that in the existing pseudo-ginseng callus culture expanding propagation technology proposed in the background technology, the supply of culture solution to the pseudo-ginseng callus is not balanced and stable enough, the whole nutrition absorption efficiency of the pseudo-ginseng callus is low, the growth period is still longer, and the rapid expanding propagation effect of the pseudo-ginseng callus cannot be achieved.
In order to achieve the above purpose, the present invention provides the following technical solutions: a culture method for expanding propagation of pseudo-ginseng green embryogenic callus comprises the following steps:
a: preparing pseudo-ginseng callus, namely selecting pseudo-ginseng green embryogenic callus induced by annual healthy pseudo-ginseng;
b: the method comprises the steps of preparing a liquid culture medium, wherein the liquid culture medium consists of four hormones of 2,4-D, 6-BA, NAA and IBA;
c: culturing green embryogenic callus of radix Notoginseng by using a paper bridge, culturing the green embryogenic callus of radix Notoginseng by using a liquid culture medium, wherein the paper bridge is a manually folded filter paper bridge, placing the filter paper bridge into the liquid culture medium, absorbing the liquid culture medium by using the paper bridge, and lifting the callus from the upper part of the paper bridge without submerging the callus in the liquid, so that the callus can quickly absorb the liquid culture medium to quickly grow.
Optionally, in the step C, the culture temperature is 20-28 ℃, and the light culture brightness is 300-500Lx.
Optionally, the mass concentration ratio of the liquid culture medium 2,4-D and 6-BA, NAA, IBA is 0.5-1.5:0.5-1.5:0.05-0.2:0.1-1.
Optionally, the liquid medium has a 2,4-D concentration of 0.5mg/L, a 6-BA concentration of 0.5mg/L, NAA concentration of 0.05mg/L, IBA concentration of 0.1mg/L.
Alternatively, the liquid medium has a 2,4-D concentration of 1.2mg/L, a 6-BA concentration of 1.2mg/L, NAA and a concentration of 0.15mg/L, IBA of 0.8mg/L.
Optionally, the liquid medium has a 2,4-D concentration of 1mg/L, a 6-BA concentration of 1mg/L, NAA concentration of 0.1mg/L, IBA concentration of 0.5mg/L.
Alternatively, the liquid medium is 1/2MS medium supplemented with 2,4-D, 6-BA, NAA and IBA.
Optionally, the pH of the liquid medium is 5-7.
Compared with the prior art, the invention provides a culture method for expanding propagation of pseudo-ginseng green embryogenic callus, which has the following beneficial effects:
1. according to the culture method for expanding propagation of the pseudo-ginseng green embryogenic callus, the liquid culture medium prepared by the formula is matched with the paper bridge for culturing the pseudo-ginseng callus, and the pseudo-ginseng callus can quickly and continuously absorb the culture solution in the liquid culture medium by the culture method, so that the nutrient absorption of the pseudo-ginseng callus is stable, balanced and efficient, and the effect of quickly expanding propagation of the pseudo-ginseng callus is achieved.
2. According to the culture method for expanding propagation of pseudo-ginseng green embryogenic callus, the liquid culture medium is composed of four hormones of 2,4-D, 6-BA, NAA and IBA, so that the pseudo-ginseng callus is higher in growth stability, high in rising speed, healthy in callus and better in growth situation.
Drawings
FIG. 1 is a photograph showing the propagation (2 weeks) of green embryogenic callus of Panax notoginseng in solid medium;
FIG. 2 is a photograph showing the propagation (2 weeks) of green embryogenic callus of Panax notoginseng (Burk.) F.H.Chen in the liquid paper bridge of the present invention.
Description of the embodiments
The invention is further illustrated by the following examples. It is to be understood that the following examples are given for illustrative purposes only and are not intended to limit the scope of the present invention. Various modifications and alterations of this invention may be made by those skilled in the art without departing from the spirit and scope of this invention.
The experimental methods used in the following examples are conventional methods unless otherwise specified.
Examples
A culture method for expanding propagation of pseudo-ginseng green embryogenic callus comprises the following steps:
a: preparing pseudo-ginseng callus, namely selecting pseudo-ginseng green embryogenic callus induced by annual healthy pseudo-ginseng;
b: preparing a liquid culture medium, wherein the liquid culture medium consists of four hormones of 2,4-D, 6-BA, NAA and IBA, the concentration of the 2,4-D is 0.5mg/L, the concentration of the 6-BA is 0.5mg/L, NAA, the concentration of the 6-BA is 0.05mg/L, IBA, the concentration of the liquid culture medium is 0.1mg/L, and the PH of the liquid culture medium is 5;
c: culturing green embryogenic callus of radix Notoginseng by using a paper bridge, wherein the paper bridge is a manually folded filter paper bridge, placing the filter paper bridge into the liquid culture medium, absorbing the liquid culture medium by using the paper bridge, and lifting the callus from the upper part of the paper bridge without submerging the callus in the liquid, so that the callus can quickly absorb the liquid culture medium to grow, and the culture temperature is 20 ℃ and the weak light culture brightness is 300Lx.
Examples
A culture method for expanding propagation of pseudo-ginseng green embryogenic callus comprises the following steps:
a: preparing pseudo-ginseng callus, and selecting green embryogenic callus induced by annual healthy pseudo-ginseng;
b: preparing a liquid culture medium, wherein the liquid culture medium consists of four hormones of 2,4-D, 6-BA, NAA and IBA, the concentration of the 2,4-D is 1.2mg/L, the concentration of the 6-BA is 1.2mg/L, NAA, the concentration of the 6-BA is 0.15mg/L, IBA, the concentration of the liquid culture medium is 0.8mg/L, and the PH of the liquid culture medium is 7;
c: culturing green embryogenic callus of radix Notoginseng by using a paper bridge, wherein the paper bridge is a manually folded filter paper bridge, placing the filter paper bridge into the liquid culture medium, absorbing the liquid culture medium by using the paper bridge, and lifting the callus from the upper part of the paper bridge without submerging the callus in the liquid, so that the callus can quickly absorb the liquid culture medium to grow, the culture temperature is 28 ℃, and the weak light culture brightness is 400Lx.
Examples
A culture method for expanding propagation of pseudo-ginseng green embryogenic callus comprises the following steps:
a: preparing pseudo-ginseng callus, and selecting green embryogenic callus induced by annual healthy pseudo-ginseng;
b: the method comprises the steps of preparing a liquid culture medium, wherein the liquid culture medium consists of four hormones of 2,4-D, 6-BA, NAA and IBA, the concentration of the 2,4-D is 1mg/L, the concentration of the 6-BA is 1mg/L, NAA, the concentration of the 6-BA is 0.1mg/L, IBA, the concentration of the liquid culture medium is 0.5mg/L, and the PH of the liquid culture medium is 5.8;
c: culturing green embryogenic callus of radix Notoginseng by using a paper bridge, wherein the paper bridge is a manually folded filter paper bridge, placing the filter paper bridge into the liquid culture medium, absorbing the liquid culture medium by using the paper bridge, and lifting the callus from the upper part of the paper bridge without submerging the callus in the liquid, so that the callus can quickly absorb the liquid culture medium to grow, and the culture temperature is 24 ℃ and the weak light culture brightness is 500Lx.
Table 1 shows hormone ratios of liquid culture media of examples 1 to 3
Table 2 shows the effect of examples 1 to 3 on the amount of cell growth
Through the three groups of embodiments, the green embryogenic callus of the pseudo-ginseng can be propagated, wherein the propagation effect of the pseudo-ginseng callus of the third group of embodiments is the best.
In the implementation of the invention, the expanding propagation culture of the pseudo-ginseng is taken as an example, and a person skilled in the art can know that the expanding propagation mode of the pseudo-ginseng is not limited to the expanding propagation of the pseudo-ginseng.
Claims (2)
1. The culture method for expanding propagation of pseudo-ginseng green embryogenic callus is characterized by comprising the following steps of:
A. preparing pseudo-ginseng callus, namely selecting pseudo-ginseng green embryogenic callus induced by annual healthy pseudo-ginseng;
B. the method comprises the steps of preparing a liquid culture medium, wherein the liquid culture medium consists of four hormones of 2,4-D, 6-BA, NAA and IBA;
C. culturing green embryogenic callus of radix Notoginseng by using a paper bridge, wherein the paper bridge is a manually folded filter paper bridge, placing the filter paper bridge into a liquid culture medium, absorbing the liquid culture medium by using the paper bridge, and lifting a callus shed on the upper part of the paper bridge without submerging the callus in the liquid, so that the callus can quickly absorb the liquid culture medium to quickly grow;
in the step C, the culture temperature is 20-28 ℃, and the weak light culture brightness is 300-500Lx;
the ratio of the four hormones in the liquid culture medium is as follows: 2,4-D concentration is 0.5mg/L, 6-BA concentration is 0.5mg/L, NAA concentration is 0.05mg/L, IBA concentration is 0.1mg/L; or 2,4-D concentration is 1.2mg/L, 6-BA concentration is 1.2mg/L, NAA concentration is 0.15mg/L, IBA concentration is 0.8mg/L; or 2,4-D at 1mg/L, 6-BA at 1mg/L, NAA at 0.1mg/L, IBA at 0.5mg/L.
2. The culture method for expanding propagation of pseudo-ginseng green embryogenic callus according to claim 1, which is characterized in that: the pH of the liquid culture medium is 5-7.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103923874A (en) * | 2014-03-25 | 2014-07-16 | 天津大学 | Pseudo-ginseng cell suspension culture method |
| CN109329056A (en) * | 2018-10-23 | 2019-02-15 | 大连工业大学 | A kind of abductive approach of Radix Notoginseng adventitious root |
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| KR101063842B1 (en) * | 2009-05-13 | 2011-09-08 | 한방바이오 주식회사 | Mass production method of wild ginseng cultured root by crystallization |
| CN102907318B (en) * | 2011-08-01 | 2015-09-09 | 东北林业大学 | A kind of method utilizing the fast numerous pseudo-ginseng regeneration plant of bioreactor culture somatic embryo |
| CN104686340A (en) * | 2015-02-22 | 2015-06-10 | 杨业容 | Method for establishing cell suspension culture system of gynura divaricate (L.) DC. |
| CN104920214B (en) * | 2015-06-03 | 2017-06-20 | 中国科学院昆明植物研究所 | A kind of method for improving pseudo-ginseng tissue-cultured seedling saponin(e yield |
| CN105993940B (en) * | 2016-05-16 | 2018-01-05 | 云南农业大学 | A kind of method grown using Induced by Salicylic Acid pseudo-ginseng stem or Callus of Leaf |
| CN106069791B (en) * | 2016-08-25 | 2018-04-24 | 文山苗乡三七科技有限公司 | A kind of pseudo-ginseng embryonic callus induction cultural method |
| CN108094203B (en) * | 2017-12-22 | 2020-11-24 | 中国农业科学院特产研究所 | Preparation method of pseudo-ginseng seeds |
| CN108849519A (en) * | 2018-08-02 | 2018-11-23 | 中国中医科学院中药研究所 | A kind of cultivating system and cultural method of fast-propagation Radix Notoginseng callus |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103923874A (en) * | 2014-03-25 | 2014-07-16 | 天津大学 | Pseudo-ginseng cell suspension culture method |
| CN109329056A (en) * | 2018-10-23 | 2019-02-15 | 大连工业大学 | A kind of abductive approach of Radix Notoginseng adventitious root |
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