CN1210663A - Seed sprouting and quick propagating technology of Yunnan Rhizoma Paridis - Google Patents
Seed sprouting and quick propagating technology of Yunnan Rhizoma Paridis Download PDFInfo
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- CN1210663A CN1210663A CN 98118955 CN98118955A CN1210663A CN 1210663 A CN1210663 A CN 1210663A CN 98118955 CN98118955 CN 98118955 CN 98118955 A CN98118955 A CN 98118955A CN 1210663 A CN1210663 A CN 1210663A
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Abstract
Yunnan Rhizoma Paridis is a main medicinal variety of Chinese medicinal materials Rhizoma Paridis. The present invention aims at solving the problems of available technology, which has slow germination speed due to rest period, and it provides a technology of germination to override the rest period and thus to propagate fast. The present invention can effectively raise the yield and improve the quality of Yunnan Rhizoma Paridis.
Description
The present invention relates to biological technical field, particularly, relate to the seed sprouting technology and the quick micro-propagation technique field of breaking the Yunnan Rhizoma Paridis seed dormancy phase.
Yunnan Rhizoma Paridis (Paris polyphylla Smith var.yunnanensis (Fr.) Hand-Mazz.) belongs to Trilliaceae (Trilliaceae) perennial herb, for the Paris flea stops a mutation of organizing (Sect.Euthyra Franch.) leafy Paris polyphylla (Paris polyphylla Smith), be the main medicinal kind of Chinese medicine Paris polyphylla.The Paris polyphylla growth is very slow, every strain year increment is about 2-3 gram fresh weight, owing to constantly find the medical value that it is new in recent years, cause demand increasing, blindly purchase and excavate in addition, cause the Paris polyphylla resource to be subjected to great destruction, Yunnan Rhizoma Paridis of using except that traditional medicine and paris polyphylla (P.polypylla var.chinensis) resource were very rare, other kind was also very few.Relying on Paris polyphylla at present is the very difficult large-scale production of Chinese patent drug of key component.And the Yunnan Rhizoma Paridis seed has the phenomenon of dormancy 1-2, and under normal circumstances germination is very poor, and asexual seedling proliferation is very slow, also unresolved this problem in the prior art.
The present invention levies problems of the prior art, providing a kind of can effectively break the Yunnan Rhizoma Paridis seed dormancy phase, successfully carry out the fast biotechnology of breeding of Yunnan Rhizoma Paridis, have that reproduction speed is fast, the cycle short, the advantage of industrial fast breeding, can improve the output and the quality of Yunnan Rhizoma Paridis simultaneously effectively.
In order to reach above-mentioned purpose of the present invention, the invention provides following technical scheme:
Yunnan Rhizoma Paridis seed sprouting and quick propagating technology thereof are got Yunnan Rhizoma Paridis (Paris polyphyllaSmith var.yunnanensis (Fr.) Hand-Mazz.) seed, and the water flushing 70% alcohol immersion 2-5 minute, is used 0.2%HgCl
2Soak the 10-30 branch, in the gibberellin of 20-80ppm concentration degree, under 22 ℃ of dark conditions, soaked 48 hours, place sterile petri dish, under 4 ℃ or 22 ℃, dark, alternately place each 1 week, carry out seed sprouting 3-5 time repeatedly, illumination every day is 10 hours then, light intensity 1000lux, with 1/2MS is minimal medium, and additional methyl (NAA) 0.5mg/L and 6-benzyl aminopurine (6BA) 0.1mg/L breed fast.
In the above-mentioned quick propagating technology, can preferably in minimal medium, add the Yunnan Rhizoma Paridis oligosaccharide.
Further specify flesh and blood of the present invention below in conjunction with content of the test of the present invention and result, the present invention's advantage compared with prior art is described simultaneously, but content of the present invention is not limited thereto.
1, cold treatment and illumination are to the influence (seeing Table 1) of seed sprouting: 3 discontinuous cold treatments (handling 1) help breaking of Yunnan Rhizoma Paridis seed dormancy phase, improve seed germination rate.Continuous 4 ℃ of low temperature (handling 4) can not improve seed germination rate.Continuous 22 ℃ of normal temperature (handling 2) can make seed sprouting, but germination rate will be lower than the germination rate of cold treatment, but illumination under the normal temperature (handling 4) is unfavorable for seed sprouting.
Table 1 cold treatment and illumination are to the influence of Yunnan Rhizoma Paridis seed sprouting
| Processing method * | The germination rate (%) of 60 days statistics | The germination rate (%) of 120 days statistics |
| ????1 ????2 ????3 ????4 | ????60 ????30 ????10 ????20 | ????90 ????50 ????20 ????30 |
*: processing method is seen embodiment 1-4.2 gibberellin treatment are to the influence of seed sprouting
Gibberellin is to carry out cold treatment has the base of obvious facilitation to seed sprouting on to the influence of Yunnan Rhizoma Paridis seed sprouting experiment.As can be seen from Table 2, (40～80ppm) handle suitable gibberellin, can reach the result similar to cold treatment at normal temperature (22 ℃) dark place bud (handling 2).If adopt the gibberellin of debita spissitudo to soak, antithetical phrase carries out discontinuous cold treatment simultaneously, can more effectively improve seed germination rate.
Consolidated statement 1 and table 2, the Yunnan Rhizoma Paridis seed is normal temperature and 4 ℃ of continuous down germinations 60 days and 120 days, and statistics is sent out rate all below 50%, illustrates that seed has the dormancy phenomenon.The dormancy that all can break the Yunnan Rhizoma Paridis kind by cold treatment and gibberellin treatment.Illumination is unfavorable for the germination of seed.
Table 2 gibberellin is to the influence of Yunnan Rhizoma Paridis seed sprouting
Table?2?Effects?of?giberellin?concentration?on?seed
germination?of?P.polyphylla?var.yunnanensis
| Gibberellin concentration C oncentration of giberellin (ppm) | Processing method * Methods of treatment | Germination rate (%) the Germination rate after 60 daya ' experiments of 60 days statistics | Germination rate (%) the Germination rate after 120 days ' experiments of 120 days statistics |
| ????0 | ????1 ????2 | ????65 ????25 | ????85 ????45 |
| ????20 | ????1 ????2 | ????65 ????45 | ????95 ????70 |
| ????40 | ????1 ????2 | ????75 ????50 | ????100 ????85 |
| ????80 | ????1 ????2 | ????70 ????55 | ????100 ????80 |
* processing method is seen embodiment 1,2.
The micropropagation of 3 Yunnan Rhizoma Paridis
The Yunnan Rhizoma Paridis seed that has germinateed is gone on the MS medium of no hormone, under illumination, cultivate, can obtain aseptic plant.Preliminary experiment is the result show, reduce inorganic salt content, help the growth of aseptic seedling, in order to help the growth of bud propagation or root, adopted a series of 1/2MS medium, kinds of culture medium that is adopted and growth of seedlings situation thereof, result's (seeing Table 3) shows, the 1/2MS medium that adds NAA0.5mg/L and 6BA0.1mg/L had both helped the propagation of bud, also helped the growth of root simultaneously.Carry out the transition to outdoorly through indoor test-tube plantlet, slowly carry out the transition to potted plantly, a potted plant week back statistics survival rate reaches 95%, and well-grown.Increased by two buds in every month by every strain test-tube plantlet and calculate, annual every strain can be bred 4096 young plants.
The different medium of table 3 influence Table 3 Effects of different media on milipropagationof P.polyphylla var.yunnanensis to the micropropagation of Yunnan Rhizoma Paridis
4 oligosaccharides are to the influence of Yunnan Rhizoma Paridis bud propagation
| Medium media | Observed result results |
| 1/2MS+NAA0.5mg/L 1/2MS+NAA0.5mg/L+6BA0.1mg/L 1/2MS+NAA0.25mg/L+6BA1.0mg/L 1/2MS+6BA1.0mg/L | The long root of plant, no bud propagation.Plant is more tiny, and bud is many, and root also growth plant is sturdy, and rhizome expands, and bud is bred less rhizome and expanded, no bud propagation |
Oligosaccharide PP-DP7, the PP-DP8 and the PP-DP9 that in medium, add variable concentrations, PRELIMINARY RESULTS (seeing Table 4) shows differentiation and the growth not significantly influence of oligosaccharide to root, but can influence the propagation of bud, the optimum concentration that PP-DP7, PP-DP8 and PP-DP9 help bud propagation is respectively 2.5ppm, 10ppm and 20ppm.
Table 4 oligosaccharide is to the influence of Yunnan Rhizoma Paridis bud propagation
Table?4?Effects?of?oligosaccharins?PP-DP7,PP-DP8?and?PP-DP9?on
shoot?multiplication?of?P.polyphylla?var.yunnanensis
+: represent that every strain test-tube plantlet increased by 1～2 bud in every month: ++: represent that every strain test-tube plantlet increased by 3 buds in every month; +++: represents that every strain test-tube plantlet increased by 4 in every month more than the bud.
| Oligosaccharide Oligosaccharin | Concentration (ppm) Concentration | Bud propagation situation Shoot multiplication |
| Contrast | ????0 | ????+ |
| ????PP-DP7 | ????2.5 ????5.0 ????10 ????20 | ????+++ ????++ ????+ ????+ |
| ????PP-DP8 | ????2.5 ????5.0 ????10 ????20 | ????+ ????++ ????+++ ????+ |
| ????PP-DP9 | ????2.5 ????5.0 ????10 ????20 | ????+ ????+ ????++ ????+++ |
From above-mentioned experimental result and conclusion excellent beneficial effect of the present invention as can be seen be:
1, adopts the quick breeding of Yunnan Rhizoma Paridis, impel the increase of rhizoma paris rhizome year increment, in a planned way develop the plant husbandry of Paris polyphylla, guarantee the supply of Paris polyphylla medicinal material, overcome the passive situation that the Paris polyphylla medicinal material relies on wild resource fully with the way of biotechnology.
2, regulate by adjustment, gibberellin, can break effectively the Yunnan Rhizoma Paridis seed dormancy phase, improved percentage of seedgermination, can reach 100%, it is very poor to have overcome the prior art germination, and the very slow shortcoming of sexual seedling proliferation.
3, have the advantage that reproduction speed is fast, the cycle is short compared to existing technology, can realize industry breeding fast.
4, help further carrying out the research and the industrialization of Yunnan Rhizoma Paridis genetic breeding (as the foundation of high yield steroid saponin content, high yield polysaccharide and oligosaccharide content strain) and Physiology and biochemistry (as relation of relation, microbiologic population and the Paris polyphylla quality of colloid and opaque Paris polyphylla etc.).
Embodiment one:
Get Yunnan Rhizoma Paridis (Paris polyphylla Smith var.yunnanensis (Fr.) Hand-Mazz.) seed, rubbing after making seed and the arillus of succulence separating, is washed in a pan out seed, dries.With the flushing of seed water, 70% alcohol immersion 2 minutes, aseptic water washing twice is used 0.2%HgCl
2Soaked 30 minutes, in the gibberellin of 40ppm concentration, under 22 ℃ of dark conditions, soaked 48 hours, being tiled on the moistening filter paper places sterile petri dish to make germinating bed, placed for 1 week down at 22 ℃ then, placed for 1 week down for 4 ℃, so hocket, after handling through 3 times, put 22 ℃ and germinate dark down.Each is treated to 100 seeds, when radicle exposes kind of skin more than 1 millimeter, promptly thinks and germinates.Be minimal medium then with 1/2MS, additional methyl (NAA) 0.5mg/L and 6-benzyl aminopurine (6BA) 0.1mg/L breed test-tube plantlet illumination every day 10 hours, light intensity 1000lux fast.
Embodiment two:
Get Yunnan Rhizoma Paridis (Paris polyphylla Smith var.yunnanensis (Fr.) Hand-Mazz.) seed, rubbing after making seed and the arillus of succulence separating, is washed in a pan out seed, dries.With the flushing of seed water, 70% alcohol immersion 5 minutes, aseptic water washing twice is used 0.2%HgCl
2Soaked 20 minutes, and in the gibberellin of 60ppm concentration, under 22 ℃ of dark conditions, soaked 48 hours, be tiled on the moistening filter paper and place sterile petri dish to make germinating bed, put 22 ℃ and germinate dark down.Each is treated to 100 seeds, when radicle exposes kind of skin more than 1 millimeter, promptly thinks and germinates.Be minimal medium then with 1/2MS, additional methyl (NAA) 0.5mg/L, 6-benzyl aminopurine (6BA) 0.1mg/L and 10ppm oligosaccharide 9 are bred test-tube plantlet illumination every day 10 hours, light intensity 1000lux fast.
Embodiment three:
Get Yunnan Rhizoma Paridis (Paris polyphylla Smith var.yunnanensis (Fr.) Hand-Mazz.) seed, rubbing after making seed and the arillus of succulence separating, is washed in a pan out seed, dries.With the flushing of seed water, 70% alcohol immersion 5 minutes, aseptic water washing is used 0.2%HgCl
2Soaked 15 minutes, and in the gibberellin of 80ppm concentration, under 22 ℃ of dark conditions, soaked 48 hours, be tiled on the moistening filter paper and place sterile petri dish to make germinating bed, put 4 ℃ and germinate dark down.Each is treated to 100 seeds, when radicle exposes kind of skin more than 1 millimeter, promptly thinks and germinates.Be minimal medium then with 1/2MS, additional methyl (NAA) 0.5mg/L, 6-benzyl aminopurine (6BA) 0.1mg/L and 20ppm oligosaccharide 8 are bred test-tube plantlet illumination every day 10 hours, light intensity 1000lux fast.
Embodiment four:
Get Yunnan Rhizoma Paridis (Paris polyphylla Smith var.yunnanensis (Fr.) Hand-Mazz.) seed, rubbing after making seed and the arillus of succulence separating, is washed in a pan out seed, dries.With the flushing of seed water, 70% alcohol immersion 3 minutes, aseptic water washing is used 0.2%HgCl
2Soaked 25 minutes, and in the gibberellin of 20ppm concentration, under 22 ℃ of dark conditions, soaked 48 hours, be tiled on the moistening filter paper and place sterile petri dish to make germinating bed, put 22 ℃ and germinate dark down.Each is treated to 100 seeds, when radicle exposes kind of skin more than 1 millimeter, promptly thinks and germinates.Be minimal medium then with 1/2MS, additional methyl (NAA) 0.5mg/L, 6-benzyl aminopurine (6BA) 0.1mg/L and 5ppm oligosaccharide 7 are bred test-tube plantlet illumination every day 10 hours, light intensity 1000lux fast.
Claims (2)
1, Yunnan Rhizoma Paridis seed sprouting and quick propagating technology thereof is characterized in that getting Yunnan Rhizoma Paridis (Parispolyphylla Smith var.yunnanensis (Fr.) Hand-Mazz.) seed, and the water flushing 70% alcohol immersion 2-5 minute, is used 0.2%HgCl
2Soak after 10-30 minute, in the gibberellin of 20-80ppm concentration, under 22 ℃ of dark conditions, soaked 48 hours, place sterile petri dish, under 4 ℃ or 22 ℃, dark, alternately place each 1 week, carry out seed sprouting 3-5 time repeatedly, illumination every day is 10 hours then, light intensity 10001ux, with 1/2MS is minimal medium, and additional methyl (NAA) 0.5mg/L and 6-benzyl aminopurine (6BA) 0.1mg/L breed fast.
2, technology according to claim 1 is characterized in that can adding the Yunnan Rhizoma Paridis oligosaccharide in minimal medium.
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| CN98118955A CN1109489C (en) | 1998-09-08 | 1998-09-08 | Seed sprouting and quick propagating technology of Yunnan Rhizoma Paridis |
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| CN98118955A CN1109489C (en) | 1998-09-08 | 1998-09-08 | Seed sprouting and quick propagating technology of Yunnan Rhizoma Paridis |
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| CN1210663A true CN1210663A (en) | 1999-03-17 |
| CN1109489C CN1109489C (en) | 2003-05-28 |
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|---|---|---|---|---|
| CN101278612B (en) * | 2008-05-20 | 2011-04-20 | 云南大学 | Method for breaking seed dormancy of Paris polyphylla var. yunnanensis |
| CN101513163B (en) * | 2009-04-03 | 2011-05-18 | 武汉大学 | Method for increasing the contents of steroid saponin in paris polyphylla rhizome by penicillium citrinum elicitor |
| CN101518184B (en) * | 2009-04-03 | 2011-07-27 | 武汉大学 | Method for improving content of various steroid saponins in rhizome of paris polyphylla var.yunnanensis |
| CN102144553A (en) * | 2011-01-25 | 2011-08-10 | 唐忠海 | Method for rapidly propagating Paris polyphylla Smith |
| CN102301956A (en) * | 2011-07-26 | 2012-01-04 | 成都大学 | Callus induction method taking paris polyphylla coleoptiles as explants |
| CN102415236A (en) * | 2011-08-19 | 2012-04-18 | 青岛中烟种子有限责任公司 | Complete nutrition formula germination accelerating method for tobacco seed |
| CN102487627A (en) * | 2011-12-09 | 2012-06-13 | 四川农业大学 | Method for effectively breaking dormancy of Paris polyphylla seeds |
| CN102835207A (en) * | 2012-09-19 | 2012-12-26 | 贵州三元太宝实业股份有限公司 | Method for reducing sleeping cycle of Paris polyphylla seeds |
| CN103053618A (en) * | 2012-12-28 | 2013-04-24 | 浙江新蓝天园林苗木科技有限公司中泰分公司 | Tree dormancy activator and preparation method thereof |
| CN103444309A (en) * | 2013-08-06 | 2013-12-18 | 中南民族大学 | Method for quickly germinating paris polyphylla var.chinensis seeds through combination of gibberellin and filter paper |
| CN103621557A (en) * | 2013-12-20 | 2014-03-12 | 贵州大学 | Special treatment agent for polyphylla seed germination |
| CN105211081A (en) * | 2015-09-11 | 2016-01-06 | 四川国光农化股份有限公司 | A kind of increase production for strong seed composition, preparation and application thereof |
| CN105325143A (en) * | 2015-10-14 | 2016-02-17 | 中国科学院昆明植物研究所 | Paris polyphylla var yunnanensis and paris polyphylla var chinensis seedling three-phase breeding technology |
| CN105685066A (en) * | 2016-03-29 | 2016-06-22 | 石棉地生中草药开发有限公司 | Rhizoma paridis seed soaking drug and preparation method thereof |
| CN106417036A (en) * | 2016-12-21 | 2017-02-22 | 西南科技大学 | Tissue culture method for inducing immature zygotic embryos of paris polyphylla to generate somatic embryos |
| CN107980634A (en) * | 2017-12-15 | 2018-05-04 | 贵州大学 | A kind of method for obtaining the suitable explant of lucid asparagus tissue-culturing rapid propagation |
| CN108307965A (en) * | 2018-03-14 | 2018-07-24 | 上海市药材有限公司 | A kind of vernalization of paris polyphylla seed and method for culturing seedlings |
| CN108811584A (en) * | 2018-06-26 | 2018-11-16 | 中国科学院成都生物研究所 | A method of promoting Paris polyphylla seed fast germination |
| CN116584384A (en) * | 2022-09-01 | 2023-08-15 | 毕节市中药研究所 | Method for effectively breaking dormancy of wild paris polyphylla seeds and inducing germination of embryo |
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| CN1016837B (en) * | 1988-11-24 | 1992-06-03 | 中国医学科学院药用植物资源开发研究所 | Accelerating germination technology for seed of american ginseng |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101278612B (en) * | 2008-05-20 | 2011-04-20 | 云南大学 | Method for breaking seed dormancy of Paris polyphylla var. yunnanensis |
| CN101513163B (en) * | 2009-04-03 | 2011-05-18 | 武汉大学 | Method for increasing the contents of steroid saponin in paris polyphylla rhizome by penicillium citrinum elicitor |
| CN101518184B (en) * | 2009-04-03 | 2011-07-27 | 武汉大学 | Method for improving content of various steroid saponins in rhizome of paris polyphylla var.yunnanensis |
| CN102144553A (en) * | 2011-01-25 | 2011-08-10 | 唐忠海 | Method for rapidly propagating Paris polyphylla Smith |
| CN102301956B (en) * | 2011-07-26 | 2012-11-14 | 成都大学 | Callus induction method taking paris polyphylla coleoptiles as explants |
| CN102301956A (en) * | 2011-07-26 | 2012-01-04 | 成都大学 | Callus induction method taking paris polyphylla coleoptiles as explants |
| CN102415236A (en) * | 2011-08-19 | 2012-04-18 | 青岛中烟种子有限责任公司 | Complete nutrition formula germination accelerating method for tobacco seed |
| CN102487627B (en) * | 2011-12-09 | 2013-11-13 | 四川农业大学 | Method for effectively breaking dormancy of Paris polyphylla seeds |
| CN102487627A (en) * | 2011-12-09 | 2012-06-13 | 四川农业大学 | Method for effectively breaking dormancy of Paris polyphylla seeds |
| CN102835207A (en) * | 2012-09-19 | 2012-12-26 | 贵州三元太宝实业股份有限公司 | Method for reducing sleeping cycle of Paris polyphylla seeds |
| CN102835207B (en) * | 2012-09-19 | 2014-01-29 | 贵州三泓药业股份有限公司 | Method for reducing sleeping cycle of Paris polyphylla seeds |
| CN103053618A (en) * | 2012-12-28 | 2013-04-24 | 浙江新蓝天园林苗木科技有限公司中泰分公司 | Tree dormancy activator and preparation method thereof |
| CN103444309A (en) * | 2013-08-06 | 2013-12-18 | 中南民族大学 | Method for quickly germinating paris polyphylla var.chinensis seeds through combination of gibberellin and filter paper |
| CN103621557A (en) * | 2013-12-20 | 2014-03-12 | 贵州大学 | Special treatment agent for polyphylla seed germination |
| CN105211081A (en) * | 2015-09-11 | 2016-01-06 | 四川国光农化股份有限公司 | A kind of increase production for strong seed composition, preparation and application thereof |
| CN105325143A (en) * | 2015-10-14 | 2016-02-17 | 中国科学院昆明植物研究所 | Paris polyphylla var yunnanensis and paris polyphylla var chinensis seedling three-phase breeding technology |
| CN105685066A (en) * | 2016-03-29 | 2016-06-22 | 石棉地生中草药开发有限公司 | Rhizoma paridis seed soaking drug and preparation method thereof |
| CN105685066B (en) * | 2016-03-29 | 2018-01-16 | 石棉地生中草药开发有限公司 | A kind of Paris polyphylla seed soaking medicament and preparation method thereof |
| CN106417036A (en) * | 2016-12-21 | 2017-02-22 | 西南科技大学 | Tissue culture method for inducing immature zygotic embryos of paris polyphylla to generate somatic embryos |
| CN107980634A (en) * | 2017-12-15 | 2018-05-04 | 贵州大学 | A kind of method for obtaining the suitable explant of lucid asparagus tissue-culturing rapid propagation |
| CN108307965A (en) * | 2018-03-14 | 2018-07-24 | 上海市药材有限公司 | A kind of vernalization of paris polyphylla seed and method for culturing seedlings |
| CN108811584A (en) * | 2018-06-26 | 2018-11-16 | 中国科学院成都生物研究所 | A method of promoting Paris polyphylla seed fast germination |
| CN116584384A (en) * | 2022-09-01 | 2023-08-15 | 毕节市中药研究所 | Method for effectively breaking dormancy of wild paris polyphylla seeds and inducing germination of embryo |
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