CN104145813A - Method for propagating stems of aristolochia fordiana - Google Patents
Method for propagating stems of aristolochia fordiana Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 35
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- 241000782501 Aristolochia fordiana Species 0.000 title abstract 5
- 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 24
- CGIDKJRJBMFXKV-UHFFFAOYSA-N 6-n'-benzylpurine-6,6-diamine Chemical compound N1=CN=C2N=CN=C2C1(N)NCC1=CC=CC=C1 CGIDKJRJBMFXKV-UHFFFAOYSA-N 0.000 claims abstract description 16
- 239000001963 growth medium Substances 0.000 claims abstract description 14
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- FAIXYKHYOGVFKA-UHFFFAOYSA-N Kinetin Natural products N=1C=NC=2N=CNC=2C=1N(C)C1=CC=CO1 FAIXYKHYOGVFKA-UHFFFAOYSA-N 0.000 claims abstract description 7
- QANMHLXAZMSUEX-UHFFFAOYSA-N kinetin Chemical compound N=1C=NC=2N=CNC=2C=1NCC1=CC=CO1 QANMHLXAZMSUEX-UHFFFAOYSA-N 0.000 claims abstract description 7
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- 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 description 6
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- YGGXZTQSGNFKPJ-UHFFFAOYSA-N methyl 2-naphthalen-1-ylacetate Chemical compound C1=CC=C2C(CC(=O)OC)=CC=CC2=C1 YGGXZTQSGNFKPJ-UHFFFAOYSA-N 0.000 claims description 6
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- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 claims description 3
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Abstract
The invention discloses a method for propagating stems of aristolochia fordiana. The method comprises the following steps: (1) preparing an explant of aristolochia fordiana; (2) culturing the explant by an induction medium, thereby obtaining test-tube plantlets after the explant is germinated, wherein the induction medium contains 0.5-2.0 mg/L of 6-benzyl amino adenine; (3) culturing the test-tube plantlets by an enrichment medium, wherein the enrichment medium contains 0.5-1.0 mg/L of kinetin, 0-0.2mg/L of 6-benzyl amino adenine and 0-0.2mg/L of indolebutyric acid; (4) culturing cluster buds of the test-tube plantlets by a strong seedling culture medium, wherein the strong seedling culture medium contains 0.2mg/L of 6-benzyl amino adenine and 0.3-0.5 mg/L of indolebutyric acid; (5) culturing strong test-tube plantlets by a rooting medium, wherein the roots of the test-tube plantlets grow by 2-4 cm, and the rooting medium contains 0.5mg/L of indolebutyric acid and 0-1.0mg/L of naphthylacetic acid; and (6) hardening seedlings, and transplanting the seedlings into a matrix for culture. According to the method for rapidly propagating stems of aristolochia fordiana, lots of excellent aristolochia fordiana seedlings suitable for transplanting can be rapidly propagated, and thus the production requirement is met.
Description
Technical field
The present invention relates to a kind of propagation method of ford dutchmanspipe root or herb stem section.
Background technology
Ford dutchmanspipe root or herb is aristolochiaceae plant, has another name called that wither in five brave Tongcheng, centering grass, natural grass,, blood rattan secretly disappears, herbaceous species, and root is cylindrical, elongated, and stem, without hair, often has vertical rib after doing; Bitter, pungent, warm in nature, slightly poisonous, there is removing toxicity for detumescence, an effect of the analgesia of dispeling the wind, promoting the circulation of qi cough-relieving, cure mainly treating rheumatic ostealgia, epigastric pain, stomachache, sphagitis, traumatic injury, child convulsion, venomous snake bite.Originate in the ground such as Guangxi (Luchuan, Cangwu, Zhaoping), Guangdong (Boluo, Deqing, spring etc.), Jiangxi (Dexing), Zhejiang and Fujian, wild ford dutchmanspipe root or herb be often born in the underbrush of mountain valley and mountain region stone gap in, require comparatively strict to growing environment, therefore Regional Distribution scope is quite limited, in recent years along with the continuous increase of market demand, collection capacity to wild resource increases gradually, and wild ford dutchmanspipe root or herb resource is on the verge of exhaustion.Utilize tissue culture rapid propagating technology, can increase fast and effectively its reproduction coefficient, can not only meet the demand of market to ford dutchmanspipe root or herb medicinal material, be conducive to again the protection of ford dutchmanspipe root or herb resource.
Summary of the invention
The present invention has designed and developed a kind of propagation method of ford dutchmanspipe root or herb stem section.In the present invention, by the method for quickly breeding of ford dutchmanspipe root or herb stem section, can go out a large amount of applicable good ford dutchmanspipe root or herb seedlings of transplanting, meet need of production by Fast-propagation.
Technical scheme provided by the invention is:
A propagation method for ford dutchmanspipe root or herb stem section, comprising:
Step (1), making ford dutchmanspipe root or herb explant;
Step (2), utilize inducing culture to cultivate explant, after explant is sprouted, obtain test-tube plantlet, in inducing culture containing the 6-benzyl aminoadenine of 0.5-2.0mg/L;
Step (3), utilize proliferated culture medium culture test tube seedling, obtain test-tube plantlet Multiple Buds, in proliferated culture medium containing the kinetin of 0.5-1.0mg/L, the 6-benzyl aminoadenine of 0-0.2mg/L, the indolebutyric acid of 0-0.2mg/L;
Step (4), utilize strong seedling culture base culture test tube seedling Multiple Buds, obtain healthy and strong test-tube plantlet, in strong seedling culture base containing the 6-benzyl aminoadenine of 0.2mg/L, the indolebutyric acid of 0.3-0.5mg/L;
Step (5), utilize root media to cultivate healthy and strong test-tube plantlet, obtain the test-tube plantlet with root, the long 2-4cm of test tube shoot root, in root media containing the indolebutyric acid of 0.5mg/L and the methyl α-naphthyl acetate of 0-1.0mg/L;
Step (6), hardening, transplant to matrix and cultivate.
Preferably, in the propagation method of described ford dutchmanspipe root or herb stem section, in described step (2), in inducing culture, contain the 6-benzyl aminoadenine of 1.5mg/L; In described step (3), in proliferated culture medium, contain the kinetin of 0.5mg/L, the 6-benzyl aminoadenine of 0.2mg/L, the indolebutyric acid of 0.2mg/L; In described step (4), in strong seedling culture base, contain the 6-benzyl aminoadenine of 0.2mg/L, the indolebutyric acid of 0.5mg/L; In described step (5), in root media, contain the indolebutyric acid of 0.5mg/L and the methyl α-naphthyl acetate of 0.6mg/L.
Preferably, in the propagation method of described ford dutchmanspipe root or herb stem section, in described step (6), matrix is mixed and is formed according to volume ratio by humus soil, vermiculite and coconut palm chaff at 3: 1: 1.
Preferably, in the propagation method of described ford dutchmanspipe root or herb stem section, described inducing culture, proliferated culture medium and strong seedling culture base all adopt MS medium, root media adopts 1/2MS medium, inducing culture, proliferated culture medium, strong seedling culture base and root media all contain agar and the 25g/L sucrose of 4.5g/L, and pH value is 5.8.
Preferably, in the propagation method of described ford dutchmanspipe root or herb stem section, in described step (1), choose on ford dutchmanspipe root or herb plant the tender stem section with terminal bud or the tender stem section with an axillalry bud as explant.
Preferably, in the propagation method of described ford dutchmanspipe root or herb stem section, in described step (1), explant is carried out disinfection, and the process of concrete sterilization is: the liquid detergent aqueous solution soaking 5-10min that is 1% by volumetric concentration, and running water rinses 15-30min, sterile water rinse once, then the alcohol-pickled 30-50s that is 75% by volumetric concentration, then the mercuric chloride solution sterilization 6-8min that is 0.1% by the mass concentration of having added 2-3 and drip Tween-20, finally use aseptic water washing 3-5 time.
Preferably, in the propagation method of described ford dutchmanspipe root or herb stem section, in described step (2), cultivation temperature is 24-26 DEG C, and intensity of illumination is 1500lux, and light application time is 10-12 hour/day, cultivates 30 days; In described step (3), cultivation temperature is 24-26 DEG C, and intensity of illumination is 2000lux, and light application time is 10-12 hour/day, cultivates 30 days; In described step (4), cultivation temperature is 24-26 DEG C, and intensity of illumination is 2000lux, and light application time is 10-12 hour/day, cultivates 21 days; In described step (5), cultivation temperature is 24-26 DEG C, and intensity of illumination is 2000lux, and light application time is 10-12 hour/day, cultivates 21 days.
Preferably, in the propagation method of described ford dutchmanspipe root or herb stem section, in described step (6), the detailed process of hardening is: the test-tube plantlet with root is shifted out to culturing room together with blake bottle, proceed to booth, be to place 7 days under the condition of 25-27 DEG C in temperature, then open bottle cap, in bottle, add running water, hardening 3-4 days.
Preferably, in the propagation method of described ford dutchmanspipe root or herb stem section, in described step (6), incubation after transplanting is specially: within after transplanting 7-10 days, use plastic film moisturizing, above plastic film, cover again one deck newspaper, shading rate is 60%-70%, and air humidity is 85%-90%, sprays every day 1 time.
The present invention carries out tissue-culturing quick-propagation by biotechnology to ford dutchmanspipe root or herb, be not subject to the restriction of season, time, cultivate at short notice in a large number and can, for the ford dutchmanspipe root or herb seedling of transplanting, significantly improve reproduction coefficient and the seedling quality of ford dutchmanspipe root or herb seedling, realized the continuity of ford dutchmanspipe root or herb provenance.Meanwhile, the present invention adopts the stem section training method with blastogenesis bud, and induction time is short, and generating process is simple, can keep preferably maternal biological character, can guarantee the medical value of ford dutchmanspipe root or herb.
Embodiment
The present invention is described in further detail below, to make those skilled in the art can implement according to this with reference to specification word.
A propagation method for ford dutchmanspipe root or herb stem section, comprises the following steps:
(1) selection of explant: get the tender stem section of ford dutchmanspipe root or herb band terminal bud or save tender stem section near the band of terminal bud, extract unnecessary blade, under flowing water, scrub gently explant surface with capillary brush, be then cut into the tender stem section with terminal bud of 2-4cm and the tender stem section with an axillalry bud, the best is 3cm.
(2) sterilization of explant: be 1% liquid detergent aqueous solution soaking 5-10min by mass concentration successively by the tender stem section of step (1), running water rinses 15-30min, sterile water rinse once, then the alcohol-pickled 30-50s that is 75% by volumetric concentration, be 0.1% mercuric chloride sterilization 7min by 150 milliliters of mass concentrations of having added 2-3 and drip Tween-20 again, aseptic water washing 3-5 time, finally spreads out on sterilized filter paper to remove surface moisture, obtains aseptic explant.
(3) aseptic bud induction: the tender stem section with terminal bud or band joint that step (2) is obtained cuts 1.0cm-2.0cm after decaptitating and truncating and is inoculated in MS medium, cultivation temperature is 24-26 DEG C, intensity of illumination 1500lux, light application time is 10-12 hour/day, cultivate 30 days, explant obtains in vitro cuttings after sprouting, wherein agar and the 25g/L sucrose of 1.5mg/L6-benzyl aminoadenine (6-benzyl aminoadenine is called for short 6-BA), 4.5g/L in MS medium, the pH value of medium is 5.8.
(4) test-tube plantlet Multiple Buds shoot proliferation is cultivated: the in vitro cuttings obtaining in step (3) is placed in to MS proliferated culture medium, temperature 24-26 DEG C, intensity of illumination 2000lux, light application time is 10-12 hour/day, cultivate and within 30 days, obtain test-tube plantlet Multiple Buds, agar and the 25g/L sucrose of 0.5mg/L kinetin (kinetin is called for short KT), 0.2mg/L6-BA, 0.2mg/L indolebutyric acid (indolebutyric acid is called for short IBA), 4.5g/L wherein in MS proliferated culture medium, are added, the pH value of medium is 5.8, KT and 6-BA promote division, and IBA promotes growth.
(5) strong seedling culture: the test-tube plantlet Multiple Buds obtaining in step (4) is placed in to MS strong seedling culture base, at cultivation temperature 24-26 DEG C, intensity of illumination 2000lux, light application time is to cultivate and within 21 days, obtain healthy and strong test-tube plantlet under the condition of 10-12 hour/day, wherein in MS strong seedling culture base, contain the powdered activated carbon of 0.5g/L, the 6-BA of 0.2mg/L, agar and the 25g/L sucrose of 0.4mg/L IBA, 4.5g/L, the pH value of medium is 5.8.6-BA is combined with IBA can favourable promotion ford dutchmanspipe root or herb test-tube plantlet growth, and along with the increase of IBA concentration, effect is better, but easy long callus of excessive concentration is advisable with 0.4mg/LIBA.
(6) culture of rootage: by the healthy and strong test-tube plantlet obtaining in step (5) be placed in 1/2MS root media (1/2MS by the macroelement of MS reduce by half other constant and configuration medium), cultivation temperature 24-26 DEG C, intensity of illumination 2000lux, light application time is 10-12 hour/day, cultivate the complete test-tube plantlet obtaining for 21 days with root, wherein in 1/2MS root media, contain the powdered activated carbon of 0.3g/L, the IBA of 0.5mg/L, 0.6mg/L methyl α-naphthyl acetate (methyl α-naphthyl acetate is called for short NAA), the agar of 4.5g/L and 25g/L sucrose, the pH value of medium is 5.8, IBA is combined with NAA and can be effectively induced ford dutchmanspipe root or herb rooting of vitro seedling.
(7) hardening and transplanting: after the whole plant with 2-4cm root length obtaining until step (6), the seedling of having taken root is shifted out to culturing room together with blake bottle, proceed to booth, temperature is 25-27 DEG C, place 7 days, then open bottle cap, in bottle, add running water, hardening 3-4 days, take out, clean root medium, be transplanted to by humus soil and vermiculite, coconut palm chaff (volume ratio is 3: 1: 1) mixes in the matrix of composition, after transplanting, within 7-10 days, use plastic film moisturizing, cover again one deck newspaper above, shading rate is that 60%-70% is the most suitable, air humidity 85%-90%, spray every day 1 time.
The present invention carries out tissue-culturing quick-propagation by biotechnology to ford dutchmanspipe root or herb, be not subject to the restriction of season, time, cultivate at short notice in a large number and can, for the ford dutchmanspipe root or herb seedling of transplanting, significantly improve reproduction coefficient and the seedling quality of ford dutchmanspipe root or herb seedling, realized the continuity of ford dutchmanspipe root or herb provenance.Meanwhile, the present invention adopts the stem section training method with blastogenesis bud, and induction time is short, and generating process is simple, can keep preferably maternal biological character, can guarantee the medical value of ford dutchmanspipe root or herb.
Although embodiment of the present invention are open as above, but it is not restricted to listed utilization in specification and embodiment, it can be applied to various applicable the field of the invention completely, for those skilled in the art, can easily realize other amendment, therefore do not deviating under the universal that claim and equivalency range limit, the present invention is not limited to specific details.
Claims (9)
1. a propagation method for ford dutchmanspipe root or herb stem section, is characterized in that, comprising:
Step (1), making ford dutchmanspipe root or herb explant;
Step (2), utilize inducing culture to cultivate explant, after explant is sprouted, obtain test-tube plantlet, in inducing culture containing the 6-benzyl aminoadenine of 0.5-2.0mg/L;
Step (3), utilize proliferated culture medium culture test tube seedling, obtain test-tube plantlet Multiple Buds, in proliferated culture medium containing the kinetin of 0.5-1.0mg/L, the 6-benzyl aminoadenine of 0-0.2mg/L, the indolebutyric acid of 0-0.2mg/L;
Step (4), utilize strong seedling culture base culture test tube seedling Multiple Buds, obtain healthy and strong test-tube plantlet, in strong seedling culture base containing the 6-benzyl aminoadenine of 0.2mg/L, the indolebutyric acid of 0.3-0.5mg/L;
Step (5), utilize root media to cultivate healthy and strong test-tube plantlet, obtain the test-tube plantlet with root, the long 2-4cm of test tube shoot root, in root media containing the indolebutyric acid of 0.5mg/L and the methyl α-naphthyl acetate of 0-1.0mg/L;
Step (6), hardening, transplant to matrix and cultivate.
2. the method for claim 1, is characterized in that, in described step (2), contains the 6-benzyl aminoadenine of 1.5mg/L in inducing culture; In described step (3), in proliferated culture medium, contain the kinetin of 0.5mg/L, the 6-benzyl aminoadenine of 0.2mg/L, the indolebutyric acid of 0.2mg/L; In described step (4), in strong seedling culture base, contain the 6-benzyl aminoadenine of 0.2mg/L, the indolebutyric acid of 0.5mg/L; In described step (5), in root media, contain the indolebutyric acid of 0.5mg/L and the methyl α-naphthyl acetate of 0.6mg/L.
3. the method for claim 1, is characterized in that, in described step (6), matrix is mixed and formed according to volume ratio by humus soil, vermiculite and coconut palm chaff at 3: 1: 1.
4. method as claimed in claim 2, it is characterized in that, inducing culture, proliferated culture medium and strong seedling culture base all adopt MS medium, root media adopts 1/2MS medium, inducing culture, proliferated culture medium, strong seedling culture base and root media all contain agar and the 25g/L sucrose of 4.5g/L, and pH value is 5.8.
5. the method as described in any one in claim 1 to 4, is characterized in that, in described step (1), chooses on ford dutchmanspipe root or herb plant the tender stem section with terminal bud or the tender stem section with an axillalry bud as explant.
6. the method as described in any one in claim 1 to 4, it is characterized in that, in described step (1), explant is carried out disinfection, and the process of concrete sterilization is: the liquid detergent aqueous solution soaking 5-10min that is 1% by volumetric concentration, and running water rinses 15-30min, sterile water rinse once, then the alcohol-pickled 30-50s that is 75% by volumetric concentration, then the mercuric chloride solution sterilization 6-8min that is 0.1% by the mass concentration of having added 2-3 and drip Tween-20, finally use aseptic water washing 3-5 time.
7. the method as described in any one in claim 1 to 4, is characterized in that, in described step (2), cultivation temperature is 24-26 DEG C, and intensity of illumination is 1500lux, and light application time is 10-12 hour/day, cultivates 30 days; In described step (3), cultivation temperature is 24-26 DEG C, and intensity of illumination is 2000lux, and light application time is 10-12 hour/day, cultivates 30 days; In described step (4), cultivation temperature is 24-26 DEG C, and intensity of illumination is 2000lux, and light application time is 10-12 hour/day, cultivates 21 days; In described step (5), cultivation temperature is 24-26 DEG C, and intensity of illumination is 2000lux, and light application time is 10-12 hour/day, cultivates 21 days.
8. the method as described in any one in claim 1 to 4, it is characterized in that, in described step (6), the detailed process of hardening is: the test-tube plantlet with root is shifted out to culturing room together with blake bottle, proceed to booth, be to place 7 days under the condition of 25-27 DEG C in temperature, then open bottle cap, in bottle, add running water, hardening 3-4 days.
9. the method as described in any one in claim 1 to 4, it is characterized in that, in described step (6), incubation after transplanting is specially: within after transplanting 7-10 days, use plastic film moisturizing, above plastic film, cover again one deck newspaper, shading rate is 60%-70%, and air humidity is 85%-90%, sprays every day 1 time.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104770296A (en) * | 2015-04-03 | 2015-07-15 | 广西壮族自治区药用植物园 | Method for rapidly propagating through adoption of aristolochia fordiana leaves |
| CN108064698A (en) * | 2018-02-05 | 2018-05-25 | 遵义市龙驰生物科技有限公司 | A kind of natural crude drugs Ciliatenerve Knotweed Root tissue culture and rapid propagation method |
| CN109247237A (en) * | 2018-11-22 | 2019-01-22 | 林登淞 | A kind of construction method of river Ciliatenerve Knotweed Root regenerating system |
| CN110742079A (en) * | 2019-10-29 | 2020-02-04 | 四川国光农化股份有限公司 | Pesticide composition for promoting root growth and increasing yield of edible leguminous crops, preparation and application |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005012507A1 (en) * | 2003-07-25 | 2005-02-10 | The University Of Melbourne | Production of plant secondary metabolites using adsorption and elicitation in cell suspension culture |
| CN103168693A (en) * | 2013-04-12 | 2013-06-26 | 广西橡胶研究所 | Proliferation culture medium for aristolochia longgonensis tissue culture and application of proliferation culture medium |
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2014
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005012507A1 (en) * | 2003-07-25 | 2005-02-10 | The University Of Melbourne | Production of plant secondary metabolites using adsorption and elicitation in cell suspension culture |
| CN103168693A (en) * | 2013-04-12 | 2013-06-26 | 广西橡胶研究所 | Proliferation culture medium for aristolochia longgonensis tissue culture and application of proliferation culture medium |
Non-Patent Citations (3)
| Title |
|---|
| BARBARA J.ET AL.: "Regeneration and plantlet development from somatic tissues of Aristolochia fimbriata", 《PLANT CELL TISSUE ORGAN CULT》 * |
| 和文佳等: "贯叶马兜铃的组织培养与快速繁殖", 《植物生理学通讯》 * |
| 邵红等: "北马兜铃组培快繁技术", 《北方园艺》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104770296A (en) * | 2015-04-03 | 2015-07-15 | 广西壮族自治区药用植物园 | Method for rapidly propagating through adoption of aristolochia fordiana leaves |
| CN108064698A (en) * | 2018-02-05 | 2018-05-25 | 遵义市龙驰生物科技有限公司 | A kind of natural crude drugs Ciliatenerve Knotweed Root tissue culture and rapid propagation method |
| CN108064698B (en) * | 2018-02-05 | 2022-08-12 | 遵义市龙驰生物科技有限公司 | Tissue culture and rapid propagation method for wild medicinal material ardisia crenata |
| CN109247237A (en) * | 2018-11-22 | 2019-01-22 | 林登淞 | A kind of construction method of river Ciliatenerve Knotweed Root regenerating system |
| CN110742079A (en) * | 2019-10-29 | 2020-02-04 | 四川国光农化股份有限公司 | Pesticide composition for promoting root growth and increasing yield of edible leguminous crops, preparation and application |
| CN110742079B (en) * | 2019-10-29 | 2021-10-08 | 四川国光农化股份有限公司 | Pesticide composition for promoting root growth and increasing yield of edible leguminous crops, preparation and application |
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| Publication number | Publication date |
|---|---|
| CN104145813B (en) | 2017-02-22 |
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Application publication date: 20141119 Assignee: Guangxi Totuweisen Agricultural Technology Co.,Ltd. Assignor: GUANGXI BOTANICAL GARDEN OF MEDICINAL PLANTS Contract record no.: X2023980045866 Denomination of invention: A Reproductive Method of Stem Segments of Tongcheng Tiger Granted publication date: 20170222 License type: Common License Record date: 20231107 |
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