LU504246B1 - A method for promoting isoflavone accumulation in Astragalus membranaceus adventitious roots - Google Patents
A method for promoting isoflavone accumulation in Astragalus membranaceus adventitious roots Download PDFInfo
- Publication number
- LU504246B1 LU504246B1 LU504246A LU504246A LU504246B1 LU 504246 B1 LU504246 B1 LU 504246B1 LU 504246 A LU504246 A LU 504246A LU 504246 A LU504246 A LU 504246A LU 504246 B1 LU504246 B1 LU 504246B1
- Authority
- LU
- Luxembourg
- Prior art keywords
- amars
- liquid medium
- ethephon
- improved
- days
- Prior art date
Links
- CJWQYWQDLBZGPD-UHFFFAOYSA-N isoflavone Natural products C1=C(OC)C(OC)=CC(OC)=C1C1=COC2=C(C=CC(C)(C)O3)C3=C(OC)C=C2C1=O CJWQYWQDLBZGPD-UHFFFAOYSA-N 0.000 title claims abstract description 31
- 235000008696 isoflavones Nutrition 0.000 title claims abstract description 31
- 241000045403 Astragalus propinquus Species 0.000 title claims abstract description 22
- 235000006533 astragalus Nutrition 0.000 title claims abstract description 20
- GOMNOOKGLZYEJT-UHFFFAOYSA-N isoflavone Chemical compound C=1OC2=CC=CC=C2C(=O)C=1C1=CC=CC=C1 GOMNOOKGLZYEJT-UHFFFAOYSA-N 0.000 title claims abstract description 18
- 238000000034 method Methods 0.000 title claims abstract description 18
- 238000009825 accumulation Methods 0.000 title claims abstract description 15
- 230000001737 promoting effect Effects 0.000 title claims abstract description 13
- 239000007788 liquid Substances 0.000 claims abstract description 38
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 36
- UDPGUMQDCGORJQ-UHFFFAOYSA-N (2-chloroethyl)phosphonic acid Chemical compound OP(O)(=O)CCCl UDPGUMQDCGORJQ-UHFFFAOYSA-N 0.000 claims abstract description 29
- 239000005976 Ethephon Substances 0.000 claims abstract description 29
- 150000002515 isoflavone derivatives Chemical class 0.000 claims abstract description 13
- 229930006000 Sucrose Natural products 0.000 claims abstract description 8
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims abstract description 8
- JTEDVYBZBROSJT-UHFFFAOYSA-N indole-3-butyric acid Chemical compound C1=CC=C2C(CCCC(=O)O)=CNC2=C1 JTEDVYBZBROSJT-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000005720 sucrose Substances 0.000 claims abstract description 8
- 238000001035 drying Methods 0.000 claims abstract description 5
- 239000012528 membrane Substances 0.000 claims description 8
- 230000036512 infertility Effects 0.000 claims description 6
- 239000002028 Biomass Substances 0.000 abstract description 3
- 239000000411 inducer Substances 0.000 abstract description 2
- 238000009776 industrial production Methods 0.000 abstract description 2
- 238000004519 manufacturing process Methods 0.000 abstract description 2
- 238000012258 culturing Methods 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 15
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 12
- ZZAJQOPSWWVMBI-UHFFFAOYSA-N calycosin Chemical compound C1=C(O)C(OC)=CC=C1C1=COC2=CC(O)=CC=C2C1=O ZZAJQOPSWWVMBI-UHFFFAOYSA-N 0.000 description 8
- HKQYGTCOTHHOMP-UHFFFAOYSA-N formononetin Chemical compound C1=CC(OC)=CC=C1C1=COC2=CC(O)=CC=C2C1=O HKQYGTCOTHHOMP-UHFFFAOYSA-N 0.000 description 8
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- WACBUPFEGWUGPB-MIUGBVLSSA-N calycosin-7-O-beta-D-glucoside Chemical compound C1=C(O)C(OC)=CC=C1C1=COC2=CC(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)=CC=C2C1=O WACBUPFEGWUGPB-MIUGBVLSSA-N 0.000 description 6
- 239000008399 tap water Substances 0.000 description 5
- 235000020679 tap water Nutrition 0.000 description 5
- QUQPHWDTPGMPEX-UHFFFAOYSA-N Hesperidine Natural products C1=C(O)C(OC)=CC=C1C1OC2=CC(OC3C(C(O)C(O)C(COC4C(C(O)C(O)C(C)O4)O)O3)O)=CC(O)=C2C(=O)C1 QUQPHWDTPGMPEX-UHFFFAOYSA-N 0.000 description 4
- MGJLSBDCWOSMHL-WFMNFSIZSA-N Ononin Natural products O(C)c1ccc(C=2C(=O)c3c(OC=2)cc(O[C@H]2[C@@H](O)[C@@H](O)[C@H](O)[C@H](CO)O2)cc3)cc1 MGJLSBDCWOSMHL-WFMNFSIZSA-N 0.000 description 4
- RIKPNWPEMPODJD-UHFFFAOYSA-N formononetin Natural products C1=CC(OC)=CC=C1C1=COC2=CC=CC=C2C1=O RIKPNWPEMPODJD-UHFFFAOYSA-N 0.000 description 4
- MGJLSBDCWOSMHL-MIUGBVLSSA-N ononin Chemical compound C1=CC(OC)=CC=C1C1=COC2=CC(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)=CC=C2C1=O MGJLSBDCWOSMHL-MIUGBVLSSA-N 0.000 description 4
- MGJLSBDCWOSMHL-UHFFFAOYSA-N ononoside Natural products C1=CC(OC)=CC=C1C1=COC2=CC(OC3C(C(O)C(O)C(CO)O3)O)=CC=C2C1=O MGJLSBDCWOSMHL-UHFFFAOYSA-N 0.000 description 4
- 239000012085 test solution Substances 0.000 description 4
- 230000000052 comparative effect Effects 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 238000010992 reflux Methods 0.000 description 2
- 241000238631 Hexapoda Species 0.000 description 1
- 206010020649 Hyperkeratosis Diseases 0.000 description 1
- 241000209504 Poaceae Species 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000001166 anti-perspirative effect Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 239000003213 antiperspirant Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000002218 hypoglycaemic effect Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 230000000324 neuroprotective effect Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000002390 rotary evaporation Methods 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
Classifications
-
- 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
- A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
- A01H6/54—Leguminosae or Fabaceae, e.g. soybean, alfalfa or peanut
-
- 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
- A01H4/002—Culture media for tissue culture
-
- 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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
-
- 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
- A01H5/00—Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy
- A01H5/06—Roots
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Cell Biology (AREA)
- Physiology (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Saccharide Compounds (AREA)
Abstract
The invention provides a method for promoting the accumulation of isoflavones in Astragalus membranaceus adventitious roots (AMARs), including the following steps: S1. inoculating AMARs into improved liquid medium B5 for 27 to 33 days; the formula of improved B5 liquid medium is as follows: 3/4B5 liquid medium + sucrose 25 ~ 35g/L+ indolebutyric acid 1.5 ~ 4.0mg/L, pH5.8 ~ 6.0; adding ethephon ethanol solution into S1 medium, the concentration of ethephon in the medium is 5-50 μM/L, and culturing for 2-7 days; S3. collecting and drying AMARs. The invention can effectively promote the increase of biomass and isoflavone content in AMARs by cultivating AMARs in improved B5 medium and adding ethephon as inducer. It is suitable for the mass production of AMARs and the industrial production of A. membranaceus isoflavones, and has high practical significance.
Description
A method for promoting isoflavone accumulation in Astragalus membranaceus adventitious roots
The invention relates to the field of biotechnology, in particular to a method for promoting the accumulation of isoflavones in Astragalus membranaceus adventitious roots (AMARSs).
Background Technology
Astragalus membranaceus is the dried root of the leguminous plant Astragalus membranaceus var. Mongholicus (Bge.) Hsiao or Astragalus membranaceus (Fisch.) Bge., it has the functions of replenishing Qi and raising Yang, strengthening surface and antiperspirant, detoxifying and discharging pus, improving water and reducing swelling, and building muscle, etc. It is often used in medicine, health products, food and other industries. Isoflavone is the main active ingredient of Astragalus membranaceus, which has pharmacological effects such as antioxidant, antiviral, hypoglycemic, neuroprotective, inhibiting melanin formation and improving fatigue, however, the yield of isoflavone is low. Astragalus membranaceus isoflavones mainly include calycosin-7-O-B-D-glucoside, ononin, calycosin and formononetin, among which calycosin-7-O-B-D-glucoside is one of the "marker compounds" to evaluate the quality of Astragalus membranaceus.
At present, Astragalus membranaceus mainly relies on artificial cultivation. However, cultivated Astragalus membranaceus has a long growth cycle and is often troubled by diseases, insects and grasses, and the isoflavone content is also unstable, which is often affected by environmental factors. In order to solve this problem and meet people's demand for Astragalus membranaceus isoflavones, tissue culture methods such as callus, hairy root culture and advective root culture have been tried to produce Astragalus membranaceus isoflavones.
However, there has been no report on improving the accumulation of isoflavones in AMARSs by using 3/4 modified BS medium combined with ethethon.
The purpose of the invention is to solve the deficiency of the above technology, provide a method to promote the accumulation of isoflavones in AMARSs, and effectively promote the increase the biomass and isoflavone content in AMARSs by adding induction factor ethephon into 3/4 improved BS liquid culture.
To realize the above purpose, the technical scheme adopted by the invention is:
A method for promoting isoflavone accumulation in AM ARS consists of the following steps:
S1. AMARs were inoculated into improved liquid medium BS for 27 to 33 days.
The formula of the improved BS liquid medium is as follows: 3/4BS liquid medium 4504246 sucrose 25 ~ 35g/L+ indolebutyric acid 1.5 ~4.0mg/L, pH 5.0 ~ 6.0;
S2, add ethephon ethanol solution to S1's improved BS liquid medium, the concentration of ethephon in the improved BS liquid medium is 5-50 u M/L, culture for 2 to 7 days;
S3. Collect AMARS cultured in S2, clean and dry.
Preferably, the culture conditions of S1 and S2 are: temperature 252°C, no light, sterility.
Preferably, the concentration of ethephon ethanol solution in S2 is 100mM/mL, and the ethephon ethanol solution is filtered and sterilized by a 0.45 ı m filter membrane.
Preferably, the drying temperature in the S3 is 55 ~ 60°C and the drying time is 2 ~ 3 days.
Compared with the prior art, the invention has the following beneficial effects:
The invention can effectively promote the increase of biomass and isoflavone content in
AMARSs by cultivating AMARs in improved BS nutrient solution, and adding ethephon as inducer. In addition, the culture process 1s simple and the conditions are mild, and it 1s suitable for the mass production of Astragalus membranaceus and the industrial production of Astragalus membranaceus isoflavones, so as to meet people's demand for Astragalus membranaceus isoflavones.
The present invention is described in detail by means of concrete embodiments. The scope of the invention is not limited to the specific embodiments.
The 3/4B5 liquid medium used in the invention means that the total volume of the liquid medium is unchanged and the nutrient composition is reduced to the original 3/4 and 3/4BS liquid medium formula is shown in Table 1.
Table 1 is the formula of 3/4BS5 liquid medium.
Embodiment1
This embodiment provides a method for promoting isoflavone accumulation in Astragalus membranaceus adventitious roots (AMARSs), including the following steps:
S1. The white, tender and differentiated adventitious roots, i.e., well-grown AMARSs, were inoculated into the improved liquid medium B5 for 30 days at a temperature of 25 +2°C, no light and sterility;
The formula of the improved B5 liquid medium was 3/4BS liquid medium + sucrose 25g/L+ indole butyric acid 1.5mg/L, pH = 5.8;
S2, ethephon ethanol solution with a concentration of 100 mm /mL was prepared and sterilized by 0.45 u m filter membrane. Ethephon ethanol solution was added into S1's improved
B5 liquid medium until the concentration of ethephon in the medium was 10 1 M/L, the cultuk&'504246 condition was 252°C, no light and sterile for 5 days;
S3. The AMARSs cultured in S2 were collected, that is, the adventitious roots were removed from the medium, rinsed with tap water, and dried at 60°C for 2 days until the constant weight.
Embodiment 2
This embodiment provides a method for promoting isoflavone accumulation in AMARS, including the following steps:
S1. The white, tender and differentiated adventitious roots, i.e., well-grown AMARS, were inoculated into the improved liquid medium BS for 30 days at a temperature of 25+2°C, no light and sterility;
The formula of the improved B5 liquid medium was 3/4BS liquid medium + sucrose 28g/L+ indolebutyric acid 2.0mg/L, pH = 5.8;
S2, ethephon ethanol solution with a concentration of 95mM/mL was prepared and sterilized by 0.45um filter membrane. Ethephon ethanol solution was added into S1's improved
BS liquid medium until the concentration of ethephon in the medium was 8 uM/L. The culture condition was 25+2°C, no light and sterile for 7 days;
S3. The AMARSs cultured in S2 were collected, that is, the AMARs were removed from the medium, rinsed with tap water, and dried at 58°C for 3 days until constant weight.
Embodiment 3
A method for promoting isoflavone accumulation in AM ARS consists of the following steps:
S1. The white, tender and differentiated AMARs were inoculated into the improved BS liquid medium for 27 days at a temperature of 25+2°C, without light and sterile;
The formula of improved BS liquid medium was 3/4B5 liquid medium + sucrose 30g/L+ indolebutyric acid 2.5mg/L, pH = 5.8;
S2, ethephon ethanol solution with a concentration of 100 mm /mL was prepared and sterilized by 0.45um filter membrane. Ethephon ethanol solution was added into S1's improved
BS liquid medium until the concentration of ethephon in the medium was 10 uM/L. The culture condition was 25+2°C, no light and sterile for 5 days;
S3. The AMARSs cultured in S2 were collected, that is, the AMARs were removed from the medium, rinsed with tap water, and dried at 55°C for 3 days until constant weight.
Embodiment 4
A method for promoting isoflavone accumulation in AM ARS consists of the following steps:
S1. The white, tender and well-differentiated AMARs were inoculated into the improved
BS liquid medium for 33 days at a temperature of 252°C, no light and sterility;
The formula of the improved liquid medium was 3/4B5 liquid medium + sucrose 32g/1-V504246 indolebutyric acid 3.0mg/L, pH = 5.8;
S2, ethephon ethanol solution with a concentration of 100mM/mL was prepared and sterilized by 0.45um filter membrane. Ethephon ethanol solution was added into S1's improved
BS liquid medium until the concentration of ethephon in the medium was SuM/L. The culture condition was 25+2°C, no light and sterile for 5 days;
S3. The AMARSs cultured in S2 were collected, that is, the adventitious roots were removed from the medium, rinsed with tap water, and dried at 60°C for 2 days until constant weight.
Embodiment 5
A method for promoting isoflavone accumulation in AM ARs of the following steps:
S1. The white, tender and well-differentiated AMARs were inoculated into the improved
BS liquid medium for 30 days at a temperature of 25+2°C, no light and sterility;
The formula of improved BS liquid medium was as follows: 3/4B5 liquid medium + sucrose 35g/L+ indolebutyric acid 4.0mg/L, pH = 5.8;
S2, ethephon ethanol solution with a concentration of 100mM/mL was prepared and sterilized by 0.45um filter membrane. Ethephon ethanol solution was added into S1's improved
BS liquid medium until the concentration of ethephon in the medium was SOUM/L. The culture condition was 25+2°C, no light and sterile for 2 days;
S3. The AMARSs cultured in S2 were collected, that is, the adventitious roots were removed from the medium, rinsed with tap water, and dried at 55°C for 3 days until constant weight.
Comparative sample 1
Commercially available standard products of calycosin-7-O-B-D-glucoside, ononin, calycosin and formononetin.
Comparative sample 2
The process is the same as in Embodiment 1, except that ethephon ethanol solution is not added, i.e. S2 is not carried out.
HPLC analysis of the content of Astragalus membranaceus isoflavones in Embodiment 1-
Embodiment 5 and comparative sample 1 and sample 2 was conducted, the specific process is as follows: 1. Experimental instruments and reagents
Experimental instruments: Shimadzu high performance Liquid chromatography system (LC-10ATVP Plus), binary high pressure gradient pump (LC-10ADVP), UV-visible dual wavelength detector (SPD-10AVP Plus);
Reagents: Fructus calycosin-7-O-B-D-glucoside, ononin, calycosin and formononetin wekd/504246 all purchased from Shanghai Ronghe Pharmaceutical Technology Co., LTD. Chromatographic grade methanol, chromatographic grade acetonitrile, ultra pure water, other analytical pure. 2. Experimental method 5 2.1 Chromatographic conditions
The chromatography was performed on C18 reverse column (5 u m, 4.6 X 250mm) with water as mobile phase A and acetonitrile as mobile phase B; the flow rate was 0.8mL/min; the detection wavelength was 230nm, the column temperature was 30°C, the sample volume was 20 1 L, and the binary gradient elution conditions were as follows: the concentration of B pump was 15— 55% in 0-30min, 55— 100% in 30-35min, 100— 15% in 35-40min, and 15% in 40- 45min. 2.2 Preparation of test solution
Weigh 0.5g Astragalus membranaceus adventitious roots (AMARs) powder in a round- bottomed flask, add SOmL analytical grade methanol, reflux for 4 hours in a water bath at 80°C, filter the reflux liquid, concentrate to dry at 60°C by rotary evaporation, then dissolve with 5mL chromatographic grade methanol, remove impurities through 0.22 u m microporous filter membrane, and filter into 10mL centrifugal tube, which is the test solution. That is, the test solution for Embodiment 1- Embodiment 5 and for the sample of proportion 2.
We accurately weighed Img of commercially available calycosin-7-O-B-D-glucoside, ononin, calycosin and formononetin, and added 0.1mL chromatography-grade methanol to fully dissolve them, and prepared them into mixed standard solution with a single standard concentration of 0.1mg/mL, that is, the test solution of the sample with a ratio of 1.
Table 1 Isoflavone content
Table 1 shows the isoflavone content of embodiment and the proportional sample, it can be seen from table 1 that AMARSs cultured by the method of the present invention has short culture cycle, high isoflavone content, and the content of calycosin-7-O-B-D-glucoside meets the requirements stipulated by the Pharmacopoeia of the People's Republic of China (2015) edition (= 0.020%, dry weight count).
The above is only the preferred embodiment of the invention, and it should be noted that, for ordinary technical personnel in the technical field, a number of improvements and refinements may be made without leaving the principle of the invention, and these improvements and refinements shall also be considered as the scope of protection of the invention.
Claims (3)
1. A method for promoting isoflavone accumulation in Astragalus membranaceus adventitious roots (AMARS), it is characterized by the following steps:
SI. AMARSs were inoculated into improved liquid medium B5 for 27 to 33 days; the formula of the improved BS liquid medium is: 3/4 BS liquid medium + sucrose 25 ~ 35g/L+ indolebutyric acid 1.5 ~4.0mg/L, pH 5.0 ~ 6.0; S2, add ethephon ethanol solution to S1's improved BS liquid medium, the concentration of ethephon in the improved BS liquid medium is 5-50 uM/L, culture for 2 to 7 days; the ethephon ethanol solution concentration is 95 ~ 100mM/mL, and the ethephon ethanol solution is filtered and sterilized by 0.45um filter membrane;
S3. Collect AMARS cultured in S2, clean and dry.
2. According to the method of promoting the accumulation of isoflavones in AMARS mentioned in Claim 1, the characteristics lie in that the culture conditions of S1 and S2 are as follows: temperature 25 + 2°C, no light and sterility.
3. According to a method of promoting the accumulation of isoflavones in AMARS mentioned in Claim 1, the characteristics are that the drying temperature in said S3 is 55 ~ 60°C and the drying time is 2 ~ 3 days.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| LU504246A LU504246B1 (en) | 2023-05-17 | 2023-05-17 | A method for promoting isoflavone accumulation in Astragalus membranaceus adventitious roots |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| LU504246A LU504246B1 (en) | 2023-05-17 | 2023-05-17 | A method for promoting isoflavone accumulation in Astragalus membranaceus adventitious roots |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| LU504246B1 true LU504246B1 (en) | 2023-11-30 |
Family
ID=88925402
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| LU504246A LU504246B1 (en) | 2023-05-17 | 2023-05-17 | A method for promoting isoflavone accumulation in Astragalus membranaceus adventitious roots |
Country Status (1)
| Country | Link |
|---|---|
| LU (1) | LU504246B1 (en) |
-
2023
- 2023-05-17 LU LU504246A patent/LU504246B1/en active IP Right Grant
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102168022B (en) | Endophytic fungus Penicillium ateckii from plant Chinese Umbrellaleaf rhizome and application thereof | |
| CN102224790B (en) | Culturing method of cordyceps militaris rich in effective component (schisandra chinensis) | |
| CN106265846A (en) | A kind of fermentation process improving Radix Rhodiolae oxidation resistance | |
| LU504246B1 (en) | A method for promoting isoflavone accumulation in Astragalus membranaceus adventitious roots | |
| CN111218406A (en) | Mucor circinelloides MF-8 and application thereof in improving content of taxifolin in rhizoma smilacis glabrae | |
| CN102363796B (en) | Method for producing glycyrrhetinic acid through microbial fermentation transformation | |
| CN102234677B (en) | A kind of method of Fusarium moniliforme microbe conversion panaxdiols saponin | |
| CN110100739B (en) | Method for promoting accumulation of isoflavone in adventitious roots of astragalus membranaceus | |
| CN114276966B (en) | Astragalus endophytic bacterium for producing isoflavone component and application thereof | |
| CN110923190A (en) | Method for improving flavone phenylpropanoid compounds of saussurea involucrate cell culture | |
| CN108034681B (en) | Method for producing catalpol by using suspension culture rehmannia stem cambium stem cells | |
| CN111996223A (en) | Method for obtaining biological polysaccharide of lucid ganoderma and isatis root by utilizing bidirectional fermentation technology | |
| CN114767757A (en) | Accurate fermentation method of compound Chinese herbal medicine by-product and application thereof | |
| CN108575758A (en) | A kind of culture media composition of Herba Saussureae Involueratae tissue cultures and its application | |
| CN101407767A (en) | Method for producing Chinese caterpillar fungus by fermentation | |
| CN101376653B (en) | Flavonol compound, and preparation and use thereof | |
| CN110511876B (en) | Epimedium koreanum endophyte, culture method and metabolite thereof | |
| Enciso-Rodríguez | Micropropagation of Valeriana edulis ssp. procera | |
| Neera et al. | Tannin production in cell cultures of Sapium sebiferum | |
| CN102943103B (en) | Penicillium fungus M1 and application thereof to increase of saponins yield in fermentation process of ginseng or Americginseng | |
| CN1388245A (en) | Method of raising taxol yield of south taxad | |
| CN110229128B (en) | Novel benzofuran type compound and extraction method thereof | |
| CN101182542A (en) | Method for genetic transforming scutellaria viscidula to obtain hairy roots producing baicalin by agrobacteriitm rhizogenes | |
| CN101701193B (en) | Method for preparing podophyllotoxin and/or astragaloside and special bacterial strain thereof | |
| CN104278068B (en) | Method for increasing content of hyperoside in phellinus igniarius liquid fermentation product |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| FG | Patent granted |
Effective date: 20231130 |