CN105494090A - Method for rapidly breeding hairy roots of vietnamese sophora roots - Google Patents
Method for rapidly breeding hairy roots of vietnamese sophora roots Download PDFInfo
- Publication number
- CN105494090A CN105494090A CN201510862709.7A CN201510862709A CN105494090A CN 105494090 A CN105494090 A CN 105494090A CN 201510862709 A CN201510862709 A CN 201510862709A CN 105494090 A CN105494090 A CN 105494090A
- Authority
- CN
- China
- Prior art keywords
- aseptic
- medium
- subprostrate sophora
- root
- roots
- 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.)
- Pending
Links
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
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides a method for rapidly breeding hairy roots of vietnamese sophora roots. The method comprises the following steps: (1) seeds of Sophora tonkinensis are disinfected and inoculated to a 1/2 MS medium for germination, and aseptic seedlings of Sophora tonkinensis are obtained; (2) R1601 Rhizobium rhizogenes is used for inducing rooting of aseptic cotyledons of Sophora tonkinensis; (3) division of hairy roots obtained in the step (2) is carried out, and the divided hairy roots are placed in a nutrient solution for rapid breeding and cultivation, and the breeding multiple reaches 39-55 times. The hairy roots of vietnamese sophora roots have the characteristics of fast growth speed, good genetic stability, and good and stable synthesis capability; partial metabolites are released to the medium, and the hairy roots can be used as alternative resources of industrial vietnamese sophora roots in large scale production.
Description
Technical field
The present invention relates to a Plant Biotechnology method, particularly a kind of method for quickly breeding of subprostrate sophora hair shape root.
Background technology
Subprostrate sophora, also claims root of subprostrate sophora, southern beans root, is the dry root and rhizome of leguminous plant sophora tonkinensis Gapnep (SophoratonkinensisGapnep.), is distributed in south China, maximum with Guangxi output.There is the effect such as Qinghuo Jiedu, swelling and pain relieving, retropharyngeal abscess, sore-throat, larynx numbness, swelling and aching of gum can be treated, breathe heavily that full heat is coughed, hepatitis, constipation, jaundice, diarrhea, haemorrhoids, the favus of the scalp, mange and mosquito, worm, dog bite etc.Medical industry subprostrate sophora is as raw material, a large amount of development the treatment injection of hepatitis, the tablet of abscess of throat and antineoplastic Chinese patent drug, simultaneously industrial also for extracting the composition such as matrine, oxymatrine, be widely used in the production of pharmacy and botanical pesticide.
Subprostrate sophora poor growth, distribution is fragmentary.Due to the increase of dosage and the expansion of the approach of use, the market demand of subprostrate sophora constantly increases, and the price of medicinal material also rises steadily, thus causes the uncontrolled of subprostrate sophora resource to be excavated, and wild resource is on the verge of exhaustion.Hair shape root because of having fast growth, genetic stability is good, synthesis capability is strong and stablize, to features such as release portion metabolites in medium, be the project verification carrier of production medicinal plant secondary metabolite.In order to alleviate the pressure of wild resource, reduce the use of wild resource, the approach that many enterprises start to utilize hair shape root to ferment is to produce the alternate resources of industrial secondary metabolite.
Summary of the invention
The object of this invention is to provide a kind of method for quickly breeding of subprostrate sophora hair shape root, it can breed subprostrate sophora hair shape root fast, for the alternate resources of follow-up large-scale culture manufacture subprostrate sophora.
The present invention achieves the above object by the following technical programs: a kind of method for quickly breeding of subprostrate sophora hair shape root, comprises the following steps:
(1) acquisition of aseptic seedling: get subprostrate sophora seed and be placed in 0.1v/v% mercuric chloride sterilization 10min, use aseptic water washing again 3 times, and do not add in the 1/2MS medium of any hormone with being inoculated into after sterilized filter paper removing surface moisture, be placed in culturing room and sprout, obtain subprostrate sophora aseptic seedling cotyledon, wherein add the agar of 30g/L sucrose and 3.4g/L in MS medium, the pH value of medium is 5.8;
(2) the aseptic cotyledon root of hair of R1601 agrobacterium rhizogenes induction subprostrate sophora: the aseptic seedling cotyledon of subprostrate sophora is put into R1601 bacterium liquid and soak 16min taking-up, take sterile water as contrast, then the bacterium liquid of explant excess surface is drawn with aseptic filter paper, be transferred on the 1/2MS solid culture medium containing acetosyringone 100 μm of ol/L and cultivate 3d acquisition hair shape root, process 30 bottles, every bottle of 1 aseptic cotyledon.When the root induced grows to about 2cm, aseptically cut, on the 1/2MS solid plate medium containing Cefotaxime Sodium 300m/L, carry out degerming cultivation, every 5d transfers 1 time, until aseptic, obtains without pili shape root.
(3) subprostrate sophora hair shape root propagation: hair shape root plant division step (2) obtained is placed in 1/2MS culture fluid and carries out Fast-propagation, cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is 8-10 hour/day, wherein 1/2MS breeds containing the benzyladenine 6-BA of methyl α-naphthyl acetate NAA, 0.1-1.0mg/L of heteroauxin IAA, 0.1-1.0mg/L of 0.1-1.0mg/L and 30g/L sucrose in culture fluid, and the pH value of medium is 5.8.
Outstanding advantages of the present invention is:
(1) this method is adopted to carry out the root propagation of subprostrate sophora hair shape, there is fast growth, genetic stability is good, synthesis capability is strong and stable, to features such as release portion metabolites in medium, breeding rate reaches 39-55 doubly, is to produce the desirable carrier of secondary metabolite.
(2) the hair shape root that this method obtains is easy to carry out amount reproduction cultivation, large-scale production can be carried out using the alternate resources as industrial subprostrate sophora to extract the secondary metabolite such as matrine, oxymatrine, meet industrial needs, there is major application prospect.
Embodiment
Below in conjunction with embodiment, technical scheme of the present invention is further illustrated.
Embodiment 1
An example of the method for quickly breeding of subprostrate sophora hair shape root of the present invention, comprises the following steps:
(1) acquisition of aseptic seedling: get subprostrate sophora seed and be placed in 0.1v/v% mercuric chloride sterilization 10min, use aseptic water washing again 3 times, and do not add in the 1/2MS medium of any hormone with being inoculated into after sterilized filter paper removing surface moisture, be placed in culturing room and sprout, obtain subprostrate sophora aseptic seedling, wherein add the agar of 30g/L sucrose and 3.4g/L in MS medium, the pH value of medium is 5.8;
(2) the aseptic cotyledon root of hair of R1601 agrobacterium rhizogenes induction subprostrate sophora: the aseptic seedling cotyledon of subprostrate sophora is put into R1601 bacterium liquid and soak 16min taking-up, take sterile water as contrast, then the bacterium liquid of explant excess surface is drawn with aseptic filter paper, be transferred on the 1/2MS solid culture medium containing acetosyringone 100 μm of ol/L and cultivate 3d acquisition hair shape root, process 30 bottles, every bottle of 1 aseptic cotyledon.When the root induced grows to about 2cm, aseptically cut, on the 1/2MS solid plate medium containing Cefotaxime Sodium 300m/L, carry out degerming cultivation, every 5d transfers 1 time, until aseptic, obtains without pili shape root.
(3) subprostrate sophora hair shape root propagation: hair shape root plant division step (2) obtained is placed in 1/2MS culture fluid and carries out Fast-propagation, cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is 8-10 hour/day, wherein 1/2MS breeds in culture fluid containing the benzyladenine 6-BA of methyl α-naphthyl acetate NAA, 0.1mg/L of heteroauxin IAA, 0.1mg/L of 0.1mg/L and 30g/L sucrose, the pH value of medium is 5.8, and the breeding rate of hair shape root is 39 times.
Embodiment 2
Another example of the method for quickly breeding of subprostrate sophora hair shape root of the present invention, comprises the following steps:
(1) acquisition of aseptic seedling: get subprostrate sophora seed and be placed in 0.1v/v% mercuric chloride sterilization 10min, use aseptic water washing again 3 times, and do not add in the 1/2MS medium of any hormone with being inoculated into after sterilized filter paper removing surface moisture, be placed in culturing room and sprout, obtain subprostrate sophora aseptic seedling cotyledon, wherein add the agar of 30g/L sucrose and 3.4g/L in MS medium, the pH value of medium is 5.8;
(2) the aseptic cotyledon root of hair of R1601 agrobacterium rhizogenes induction subprostrate sophora: the aseptic seedling cotyledon of subprostrate sophora is put into R1601 bacterium liquid and soak 16min taking-up, take sterile water as contrast, then the bacterium liquid of explant excess surface is drawn with aseptic filter paper, be transferred on the 1/2MS solid culture medium containing acetosyringone 100 μm of ol/L and cultivate 3d acquisition hair shape root, process 30 bottles, every bottle of 1 aseptic cotyledon.When the root induced grows to about 2cm, aseptically cut, on the 1/2MS solid plate medium containing Cefotaxime Sodium 300m/L, carry out degerming cultivation, every 5d transfers 1 time, until aseptic, obtains without pili shape root;
(3) subprostrate sophora hair shape root propagation: hair shape root plant division step (2) obtained is placed in 1/2MS culture fluid and carries out Fast-propagation, cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is 8-10 hour/day, wherein 1/2MS breeds in culture fluid containing the benzyladenine 6-BA of methyl α-naphthyl acetate NAA, 0.5mg/L of heteroauxin IAA, 0.5mg/L of 0.8mg/L and 30g/L sucrose, the pH value of medium is 5.8, and the breeding rate of hair shape root is 55 times.
Embodiment 3
An example of the method for quickly breeding of subprostrate sophora hair shape root of the present invention, comprises the following steps:
(1) acquisition of aseptic seedling: get subprostrate sophora seed and be placed in 0.1v/v% mercuric chloride sterilization 10min, use aseptic water washing again 3 times, and do not add in the 1/2MS medium of any hormone with being inoculated into after sterilized filter paper removing surface moisture, be placed in culturing room and sprout, obtain subprostrate sophora aseptic seedling cotyledon, wherein add the agar of 30g/L sucrose and 3.4g/L in MS medium, the pH value of medium is 5.8;
(2) the aseptic cotyledon root of hair of R1601 agrobacterium rhizogenes induction subprostrate sophora: the aseptic seedling cotyledon of subprostrate sophora is put into R1601 bacterium liquid and soak 16min taking-up, take sterile water as contrast, then the bacterium liquid of explant excess surface is drawn with aseptic filter paper, be transferred on the 1/2MS solid culture medium containing acetosyringone 100 μm of ol/L and cultivate 3d acquisition hair shape root, process 30 bottles, every bottle of 1 aseptic cotyledon.When the root induced grows to about 2cm, aseptically cut, on the 1/2MS solid plate medium containing Cefotaxime Sodium 300m/L, carry out degerming cultivation, every 5d transfers 1 time, until aseptic, obtains without pili shape root;
(3) subprostrate sophora hair shape root propagation: hair shape root plant division step (2) obtained is placed in 1/2MS culture fluid and carries out Fast-propagation, cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is 8-10 hour/day, wherein 1/2MS breeds in culture fluid containing the benzyladenine 6-BA of methyl α-naphthyl acetate NAA, 1.0mg/L of heteroauxin IAA, 0.5mg/L of 0.5mg/L and 30g/L sucrose, the pH value of medium is 5.8, and the breeding rate of hair shape root is 45 times.
Embodiment 4
An example of the method for quickly breeding of subprostrate sophora hair shape root of the present invention, comprises the following steps:
(1) acquisition of aseptic seedling: get subprostrate sophora seed and be placed in 0.1v/v% mercuric chloride sterilization 10min, use aseptic water washing again 3 times, and do not add in the 1/2MS medium of any hormone with being inoculated into after sterilized filter paper removing surface moisture, be placed in culturing room and sprout, obtain subprostrate sophora aseptic seedling cotyledon, wherein add the agar of 30g/L sucrose and 3.4g/L in MS medium, the pH value of medium is 5.8;
(2) the aseptic cotyledon root of hair of R1601 agrobacterium rhizogenes induction subprostrate sophora: the aseptic seedling cotyledon of subprostrate sophora is put into R1601 bacterium liquid and soak 16min taking-up, take sterile water as contrast, then the bacterium liquid of explant excess surface is drawn with aseptic filter paper, be transferred on the 1/2MS solid culture medium containing acetosyringone 100 μm of ol/L and cultivate 3d acquisition hair shape root, process 30 bottles, every bottle of 1 aseptic cotyledon.When the root induced grows to about 2cm, aseptically cut, on the 1/2MS solid plate medium containing Cefotaxime Sodium 300m/L, carry out degerming cultivation, every 5d transfers 1 time, until aseptic, obtains without pili shape root;
(3) subprostrate sophora hair shape root propagation: hair shape root plant division step (2) obtained is placed in 1/2MS culture fluid and carries out Fast-propagation, cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is 8-10 hour/day, wherein 1/2MS breeds in culture fluid containing the benzyladenine 6-BA of methyl α-naphthyl acetate NAA, 1.0mg/L of heteroauxin IAA, 1.0mg/L of 1.0mg/L and 30g/L sucrose, the pH value of medium is 5.8, and the breeding rate of hair shape root is 46 times.
Claims (1)
1. a method for quickly breeding for subprostrate sophora hair shape root, is characterized in that: the method comprises the following steps:
(1) acquisition of aseptic seedling: get subprostrate sophora seed and be placed in 0.1v/v% mercuric chloride sterilization 10min, use aseptic water washing again 3 times, and do not add in the 1/2MS medium of any hormone with being inoculated into after sterilized filter paper removing surface moisture, be placed in culturing room and sprout, obtain subprostrate sophora aseptic seedling cotyledon, wherein add the agar of 30g/L sucrose and 3.4g/L in MS medium, the pH value of medium is 5.8;
(2) the aseptic cotyledon root of hair of R1601 agrobacterium rhizogenes induction subprostrate sophora: the aseptic seedling cotyledon of subprostrate sophora is put into R1601 bacterium liquid and soak 16min taking-up, take sterile water as contrast, then the bacterium liquid of explant excess surface is drawn with aseptic filter paper, be transferred to add acetosyringone 100 μm of ol/L 1/2MS solid culture medium on cultivate 3d and obtain hair shape root, process 30 bottles, every bottle of 1 aseptic cotyledon, when the root induced grows to about 2cm, aseptically cut, degerming cultivation is carried out on the 1/2MS solid plate medium adding Cefotaxime Sodium 300m/L, every 5d transfers 1 time, until aseptic, obtain without pili shape root,
(3) subprostrate sophora hair shape root propagation: hair shape root plant division step (2) obtained is placed in 1/2MS breeding culture fluid and carries out Fast-propagation, cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is 8-10 hour/day, wherein 1/2MS breeds the benzyladenine 6-BA and the 30g/L sucrose that add methyl α-naphthyl acetate NAA, 0.1-1.0mg/L of heteroauxin IAA, 0.1-1.0mg/L of 0.1-1.0mg/L in culture fluid, and the pH value of medium is 5.8.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510862709.7A CN105494090A (en) | 2015-11-30 | 2015-11-30 | Method for rapidly breeding hairy roots of vietnamese sophora roots |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510862709.7A CN105494090A (en) | 2015-11-30 | 2015-11-30 | Method for rapidly breeding hairy roots of vietnamese sophora roots |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105494090A true CN105494090A (en) | 2016-04-20 |
Family
ID=55702707
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510862709.7A Pending CN105494090A (en) | 2015-11-30 | 2015-11-30 | Method for rapidly breeding hairy roots of vietnamese sophora roots |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105494090A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106508675A (en) * | 2016-10-27 | 2017-03-22 | 潘克稳 | Seed tissue culture medium for root of subprostrate sophora |
| CN107624534A (en) * | 2017-10-17 | 2018-01-26 | 伊犁师范学院 | A kind of method that Sophora alopecuroide seedling is efficiently bred using rhizome |
| CN112005884A (en) * | 2020-09-14 | 2020-12-01 | 广西壮族自治区药用植物园 | Method for improving alkaloid content of subprostrate sophora tissue culture seedling by utilizing selenium |
| CN112154914A (en) * | 2020-10-09 | 2021-01-01 | 广西壮族自治区药用植物园 | Method for improving alkaloid content of subprostrate sophora tissue culture seedling by using calcium ions |
| CN115299346A (en) * | 2022-09-06 | 2022-11-08 | 广西壮族自治区药用植物园 | Method for improving alkaloid and flavone contents of subprostrate sophora tissue culture seedlings by using magnesium |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102301951A (en) * | 2011-07-18 | 2012-01-04 | 广西壮族自治区药用植物园 | Method for rapidly propagating roots of subprostrate sophora by tissue culture |
| CN104770298A (en) * | 2015-04-13 | 2015-07-15 | 广西壮族自治区药用植物园 | Method for inducing hairy roots of sophora tonkinensis roots |
-
2015
- 2015-11-30 CN CN201510862709.7A patent/CN105494090A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102301951A (en) * | 2011-07-18 | 2012-01-04 | 广西壮族自治区药用植物园 | Method for rapidly propagating roots of subprostrate sophora by tissue culture |
| CN104770298A (en) * | 2015-04-13 | 2015-07-15 | 广西壮族自治区药用植物园 | Method for inducing hairy roots of sophora tonkinensis roots |
Non-Patent Citations (2)
| Title |
|---|
| 李林轩等: ""正交试验优化山豆根组织培养条件"", 《中药材》 * |
| 王冲之等: ""不同培养基及外源激素对西洋参毛状根的生长和皂甙含量的影响"", 《植物资源与环境学报》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106508675A (en) * | 2016-10-27 | 2017-03-22 | 潘克稳 | Seed tissue culture medium for root of subprostrate sophora |
| CN107624534A (en) * | 2017-10-17 | 2018-01-26 | 伊犁师范学院 | A kind of method that Sophora alopecuroide seedling is efficiently bred using rhizome |
| CN112005884A (en) * | 2020-09-14 | 2020-12-01 | 广西壮族自治区药用植物园 | Method for improving alkaloid content of subprostrate sophora tissue culture seedling by utilizing selenium |
| CN112005884B (en) * | 2020-09-14 | 2022-02-22 | 广西壮族自治区药用植物园 | Method for improving alkaloid content of subprostrate sophora tissue culture seedling by utilizing selenium |
| CN112154914A (en) * | 2020-10-09 | 2021-01-01 | 广西壮族自治区药用植物园 | Method for improving alkaloid content of subprostrate sophora tissue culture seedling by using calcium ions |
| CN115299346A (en) * | 2022-09-06 | 2022-11-08 | 广西壮族自治区药用植物园 | Method for improving alkaloid and flavone contents of subprostrate sophora tissue culture seedlings by using magnesium |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105494090A (en) | Method for rapidly breeding hairy roots of vietnamese sophora roots | |
| CN102422811B (en) | Dendrobium officinale tissue culture reproduction method | |
| CN102301951B (en) | Method for rapidly propagating roots of subprostrate sophora by tissue culture | |
| CN103828715A (en) | Bletilla striata tissue culture breeding method | |
| CN103535281B (en) | Tissue culture medium of sealwort roots and stems and in-vitro regeneration method | |
| CN101869591B (en) | Method for producing alkannin by utilizing Arnebia euchroma(Royle)Johnst hairy root | |
| CN107223564B (en) | A kind of root of purple-flowered peucedanum plant regeneration system method for building up | |
| CN104067939A (en) | Tissue culture rapid propagation method of radix gentianae | |
| CN105493861A (en) | Method for directly seeding bletilla striata protocorms | |
| CN103270946B (en) | A kind of seedlings of Dendrobium officinale is produced and hardening synchronous method | |
| CN105165626A (en) | Culture medium formula of plant callus induction and subculture multiplication of astragalus membranaceus (fisch.) bunge and culture method of astragalus membranaceus (fisch.) bunge | |
| CN104770298B (en) | A kind of abductive approach of Radix Sophorae Tonkinensis hairy root | |
| CN101953300B (en) | Tissue culture method for Curcuma wenyujin No.1 | |
| CN103798137A (en) | Tissue culture rapid propagation method of salvia miltiorrhiza bge | |
| CN110604049B (en) | Wild-returning ecological planting method for dendrobium officinale | |
| CN105284625A (en) | Novel seedling cultivating method of dendrobium candidum | |
| CN103477976A (en) | Stem tissue culture seedling method of dendrobium candidum | |
| CN105766638A (en) | Induction method and application of dryopteris fragrans callus tissues | |
| CN109526745A (en) | A method of seedling is bred with paris polyphylla blade | |
| CN105746344A (en) | Virus-free tissue culture and rapid propagation method of castor oil plants | |
| CN104145817A (en) | Tissue culture rapid propagation technology for ashitaba leaves | |
| CN102283124B (en) | Isatis indigotica Fort. root explant organogenesis method | |
| CN106942052B (en) | A kind of industrial seedling rearing and artificial cultivation method of dryopteris fragrans | |
| CN104255472A (en) | Method for quickly reproducing semen cassiae sophera | |
| CN104472360A (en) | Method for rejuvenating dendrobium officinale tissue culture thin and weak seedlings |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20160420 |
|
| WD01 | Invention patent application deemed withdrawn after publication |