CN108566888A - A kind of medicinal plant jatamans valeriana rhizome tissue culture mating system - Google Patents
A kind of medicinal plant jatamans valeriana rhizome tissue culture mating system Download PDFInfo
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- CN108566888A CN108566888A CN201711183381.1A CN201711183381A CN108566888A CN 108566888 A CN108566888 A CN 108566888A CN 201711183381 A CN201711183381 A CN 201711183381A CN 108566888 A CN108566888 A CN 108566888A
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- jatamans valeriana
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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Abstract
The invention discloses a kind of medicinal plant jatamans valeriana rhizome tissue culture mating systems, include the following steps：S1, selection：It chooses and grows excellent and healthy and strong wild jatamans valeriana rhizome, it is positioned under tap water and rinses 1 hour or more, then it fully submerges explant in the alcohol with 75% on desinfection chamber superclean bench and handles 30s, then aseptic water washing is used 4~5 times, 10~15min is fully impregnated with 0.1% mercuric chloride solution again, is finally rinsed 5~6 times, is then shaken repeatedly thoroughly to dispose mercury chloride repeatedly with sterile water, it is spare after blotting extra moisture with aseptic filter paper again；The medicinal plant jatamans valeriana rhizome tissue culture mating system, incubation step is simple, it is easy to operate, breeding convenient for staff to plant jatamans valeriana rhizome tissue culture, using induction of callus, and compared by multigroup experiment, top quality breeding condition is filtered out convenient for user, meets the demand of medicinal plant jatamans valeriana rhizome industrialized tissue culture breeding.
Description
Technical field
The present invention relates to medicinal plant jatamans valeriana rhizome Cultivating techniques field, specially a kind of medicinal plant jatamans valeriana rhizome tissue culture breeding
Method.
Background technology
Jatamans valeriana rhizome（Valeriana jatamansi Jones.）, it is Valerianaceae valerian, the dry root of spider grass
Stem and root, the herbaceos perennials such as alias RHIZOMA ET RADIX VALERIANAE JATNANSI, lobus cardiacus valerian, tiger seven, high 30~70cm.Stem is usually grown thickly for several,
It is close by pubescence.Root-like stock it is horizontal walk, plump, blocky, coarse, internode is close, has petiole residue, yellowish-brown to have special fragrance.It is logical
It is frequently grown in the dark and damp hayashishita in river valley, border open ground, river shoal, Lin Zhong, from the grass, mountain top grassy marshland, hillside is flush in roadside.Distribution
Major area in Shaanxi Province, Hubei Province, Hunan Province, Chongqing City, Sichuan Province, Guangxi autonomous region, Tibet Autonomous Region.Li Shizhen (1518-1593 A.D.) exists
《Compendium of Materia Medica》Middle evaluation jatamans valeriana rhizome "《Shi Zhen》It says: jatamans valeriana rhizome goes out in the mountain of Songpan Xi Maozhou, another name for Sichuan Province, grass roots.Black has thick palpus, shape
If spider and this ligusticumic, rhizome of chuanxiong are poor, fragrant odour, that person also weighs it.Or cloud cat eating it ".Jatamans valeriana rhizome surveys title " seedling medicine is classical ".
Wherein mainly contain volatile oil, iridoids, flavonoids, alkaloid, amino acid and lignanoid and some other
Chemical composition, pharmacological activity includes calmness, hypnosis, antiviral, antibacterial, antitumor, antitumor cell is malicious and to the circulatory system
Effect.《Compendium of Materia Medica》In clearly describe jatamans valeriana rhizome and can ward off pestilence, calmness, control gas clothes, dehumidifying cold dispelling, taste food
Stagnant, external application boil, the common diseases such as influenza, infantile malnutrition due to digestive disturbances or intestinalparasites, malaria have good therapeutic effect.
Nowadays, the research of jatamans valeriana rhizome has caused relevant industries to be paid attention to, and is concentrated mainly on jatamans valeriana rhizome medicinal chemistry, pharmacology is made
With, volatile ingredient, quality of medicinal material standard and quality of medicinal material analysis etc., but in terms of the Plant Tissue Breeding of jatamans valeriana rhizome temporarily
In season someone did relevant research, and for jatamans valeriana rhizome because use is wider, usage amount is larger, and wild resource is more and more poorer, supply and demand
Contradiction is increasingly prominent.Existing medicinal plant jatamans valeriana rhizome tissue culture breeding, general step is extremely complex, and right in incubation
Plant and condition of culture require it is relatively high so that the survival rate of jatamans valeriana rhizome transplanting is very low, using with limitation,
It cannot be satisfied batch production demand.
Invention content
（One）The technical issues of solution
In view of the deficiencies of the prior art, the present invention provides a kind of medicinal plant jatamans valeriana rhizome tissue culture mating system, background is solved
The problem of being previously mentioned in technology.
（Two）Technical solution
In order to achieve the above object, the present invention is achieved by the following technical programs：A kind of medicinal plant jatamans valeriana rhizome tissue culture breeding
Method includes the following steps：
S1, selection：
It chooses and grows excellent and healthy and strong wild jatamans valeriana rhizome, be positioned under tap water and rinse 1 hour or more, it is then super in desinfection chamber
Fully submerge explant processing 30s with 75% alcohol on net workbench, then with aseptic water washing 4~5 times, then with 0.1%
Mercuric chloride solution fully impregnates 10~15min, is finally rinsed 5~6 times, is then shaken repeatedly with thoroughly by chlorine repeatedly with sterile water
Change mercury to dispose, then after blotting extra moisture with aseptic filter paper, it is spare；
It is prepared by S2, culture medium：
MS minimal mediums, sucrose and agar are chosen, is separately added into culture vessel and uniformly mixes, pH value 5.6, then to culture
The plant growth regulator of various concentration and type, sterilization time 30min are added in vessel；
S3, induction of callus：
The lay-by material for choosing clean is placed on superclean bench, sterile cleaning is carried out；Then lay-by material after cleaning is chosen
Subterranean stem lateral bud (length 0.5cm), petiole (length be 1. 0cm) and blade (area 2.0cm2) it is seeded to induction training
It supports on base, 5 examination materials of every bottle of inoculation, each 30 bottles of processing inoculation, 3 repetitions, starts to observe after 7d, callus group is counted after 30d
Knit induction situation；
S4, callus proliferation culture：
The callus successfully induced is seeded on test medium, 5 examination materials of every bottle of inoculation, each processing inoculation 30
Bottle weighs every bottle of callus fresh weight, weighs again after cultivating 45d, and 3 repetitions count callus proliferative conditions, filter out jatamans valeriana rhizome and be cured
Injured tissue is proliferated optimum medium；
S5, culture of rootage：
By+0.5~2.0 mgL of well-grown jatamans valeriana rhizome tissue-cultured seedling access 1/2MS minimal mediums-1 IAA is cultivated,
Each 20 bottles of processing inoculation, 3 repetitions, 60 d are 1 period, observe and count the situation of taking root of jatamans valeriana rhizome seedling strong sprout,
Filter out the culture medium of suitable jatamans valeriana rhizome seedling strengthening seedling and rooting culture.
S6, acclimatization and transplants：
Wait for that jatamans valeriana rhizome tissue-cultured seedling grows to 4~5cm, when 3~4cm of root long, transplanting is to heliogreenhouse after 3~5 d of room temperature lower refining seedling
In；Cultivation temperature is 25 ± 2 DEG C, humidity 70%~80%.Cultivation matrix is turf：Perlite（1：1）, growing state is observed,
Statistics transplanting survival rate.
Preferably, the sterile cleaning in the S3 is to handle 30s, aseptic water washing 3 to lay-by material using 75% alcohol
~5 times, then with 0.1% HgCl210~15min is fully impregnated, sterile water rinses 4~6 times repeatedly, and fully shaking is by HgCl2Clearly
It removes, blots explant surface moisture with aseptic filter paper, and be unfolded to place.
The present invention provides a kind of medicinal plant jatamans valeriana rhizome tissue culture mating systems.Has following advantageous effect：
The medicinal plant jatamans valeriana rhizome tissue culture mating system, incubation step is simple, easy to operate, convenient for staff to plant jatamans valeriana rhizome
The breeding of tissue culture is compared using induction of callus, and by multigroup experiment, is filtered out convenient for user top quality
Breeding condition meets the demand of medicinal plant jatamans valeriana rhizome tissue culture breeding.
Specific implementation mode
The present invention provides a kind of technical solution：A kind of medicinal plant jatamans valeriana rhizome tissue culture mating system, includes the following steps：
S1, selection：
It chooses and grows excellent and healthy and strong wild jatamans valeriana rhizome, be positioned under tap water and rinse 1 hour or more, it is then super in desinfection chamber
Fully submerge explant processing 30s with 75% alcohol on net workbench, then with aseptic water washing 4~5 times, then with 0.1%
Mercuric chloride solution fully impregnates 10~15min, is finally rinsed 5~6 times, is then shaken repeatedly with thoroughly by chlorine repeatedly with sterile water
Change mercury to dispose, then after blotting extra moisture with aseptic filter paper, it is spare；
It is prepared by S2, culture medium：
MS minimal mediums, sucrose and agar are chosen, is separately added into culture vessel and uniformly mixes, pH value 5.6, then to culture
The plant growth regulator of various concentration and type, sterilization time 30min are added in vessel；
S3, induction of callus：
The lay-by material for choosing clean is placed on superclean bench, sterile cleaning is carried out；Then lay-by material after cleaning is chosen
Subterranean stem lateral bud (length 0.5cm), petiole (length be 1. 0cm) and blade (area 2.0cm2) it is seeded to induction training
It supports on base, 5 examination materials of every bottle of inoculation, each 30 bottles of processing inoculation, 3 repetitions, starts to observe after 7d, callus group is counted after 30d
Knit induction situation；Sterile cleaning is to handle 30s to lay-by material using 75% alcohol, aseptic water washing 4~5 times, then with 0.1%
HgCl210~15min is fully impregnated, sterile water rinses 4~6 times repeatedly, and fully shaking is by HgCl2It removes, is inhaled with aseptic filter paper
Dry explant surface moisture, and be unfolded to place spare.
1 healing tissue inducting test of table designs
S4, callus proliferation culture：
The callus successfully induced is seeded on test medium, as shown in table 2,5 examination materials of every bottle of inoculation, each
30 bottles of processing inoculation, weighs every bottle of callus fresh weight, weighs again after cultivating 45d, 3 repetitions, counts callus proliferative conditions, screening
Go out jatamans valeriana rhizome callus proliferation optimum medium；
2 callus proliferation experimental design of table
S5, culture of rootage：
By+0.5~2.0 mgL of well-grown jatamans valeriana rhizome tissue-cultured seedling access 1/2MS minimal mediums-1IAA is trained
It supports, each 20 bottles of processing inoculation, 3 repetitions, 60 d are 1 period, observe and count the feelings of taking root of jatamans valeriana rhizome seedling strong sprout
Condition filters out the culture medium of suitable jatamans valeriana rhizome seedling strengthening seedling and rooting culture.
S6, acclimatization and transplants：
Wait for that jatamans valeriana rhizome tissue-cultured seedling grows to 4~5cm, when 3~4cm of root long, transplanting is to heliogreenhouse after 3~5 d of room temperature lower refining seedling
In；Cultivation temperature is 25 ± 2 DEG C, humidity 70%~80%.Cultivation matrix is turf：Perlite（1：1）, growing state is observed,
Statistics transplanting survival rate.
Different parts explant evoked callus is analyzed：
Jatamans valeriana rhizome subterranean stem lateral bud culture starts to expand after 6~8d, and more callus is formed after 30d, is cultivated in A6 in the short time
It can induce out adventitious bud in base and bud number be more, growing way is good；Using petiole as explant culture, 5d or so base portions first expand, then
Ultimate swelling, petiole color are shoaled by green, and more callus, but easy browning are formed after 30d；Leaf culture is first at leaf margin in 7d
Crystalline lens is generated, 23d or so forms a large amount of callus, different degrees of browning occurs.As shown in table 3, petiole callus induction is best
Culture medium is A5, inductivity 72.7%；Blade callus induction optimal medium is A7, inductivity 91.3%；Subterranean stem lateral bud
Callus induction optimal medium is A5, and inductivity 88.7%, callus growth gesture is best, as shown in table 3.
3 different culture media of table lures explant the influence of callus induction
Note："+" shows that growing way is general, and "-" shows to grow without callus or not substantially, and " ++ " shows to grow fine.It is different small
Lowercase alphabet shows notable in 0.05 level difference.
Callus proliferation culture assays
4 callus proliferation culture of table
After callus subinoculation 45d, as shown in Table 4, the callus in B1~B4 culture mediums only differentiates a small amount of
Bud does not generate new callus；Callus in B5, B6, B7, B8 and B10 culture medium generates new be cured in 15d or so
Hinder, and differentiation and bud formation, the callus in B6 and B7 culture mediums also differentiates a little adventitious root；Cultivation effect is most preferably in B9
Callus in culture medium can not only quickly generate callus, and Bud Differentiation number is most, and growing way is best, and proliferation rate is reachable
93.3%。
Culture of rootage is analyzed
Influences of the 5 various concentration NAA of table to taking root
As shown in Table 5, the radical of jatamans valeriana rhizome test tube seedling, root long and rooting rate are increased with the increase of NAA concentration in culture medium
It is long, when concentration is more than 1. 5 mgL-1, easily take root on callus though radical continues to increase.Therefore with 1/2MS+
1. 5 mgL-1 of NAA carry out culture of rootage, and for root growth at vein, radical is more, and root growth is normal, the most suitable
Preferably, rooting rate is up to 100%.
Acclimatization and transplants
According to the test method, jatamans valeriana rhizome tissue-cultured seedling transplanting survival rate after hardening is up to 95% or more.20d or so is grown newly
Leaf and new root, tissue-cultured seedling growth potential is good, and rare lesion lodging achieves the desired results.
It should be noted that herein, relational terms such as first and second and the like are used merely to a reality
Body or operation are distinguished with another entity or operation, are deposited without necessarily requiring or implying between these entities or operation
In any actual relationship or order or sequence.Moreover, the terms "include", "comprise" or its any other variant are intended to
Non-exclusive inclusion, so that the process, method, article or equipment including a series of elements is not only wanted including those
Element, but also include other elements that are not explicitly listed, or further include for this process, method, article or equipment
Intrinsic element.In the absence of more restrictions.
It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
Understanding without departing from the principles and spirit of the present invention can carry out these embodiments a variety of variations, modification, replace
And modification, the scope of the present invention is defined by the appended.
Claims (2)
1. a kind of medicinal plant jatamans valeriana rhizome tissue culture mating system, which is characterized in that include the following steps：
S1, selection：
It chooses and grows excellent and healthy and strong wild jatamans valeriana rhizome, be positioned under tap water and rinse 1 hour or more, it is then super in desinfection chamber
Fully submerge explant processing 30s with 75% alcohol on net workbench, then with aseptic water washing 4~5 times, then with 0.1%
Mercuric chloride solution fully impregnates 10~15min, is finally rinsed 5~6 times, is then shaken repeatedly with thoroughly by chlorine repeatedly with sterile water
Change mercury to dispose, then after blotting extra moisture with aseptic filter paper, it is spare；
It is prepared by S2, culture medium：
MS minimal mediums, sucrose and agar are chosen, is separately added into culture vessel and uniformly mixes, pH value 5.6, then to culture
The plant growth regulator of various concentration and type, sterilization time 30min are added in vessel；
S3, induction of callus：
The lay-by material for choosing clean is placed on superclean bench, sterile cleaning is carried out；Then lay-by material after cleaning is chosen
Subterranean stem lateral bud (length 0.5cm), petiole (length be 1. 0cm) and blade (area 2.0cm2) it is seeded to induction training
It supports on base, 5 examination materials of every bottle of inoculation, each 30 bottles of processing inoculation, 3 repetitions, starts to observe after 7d, callus group is counted after 30d
Knit induction situation；
S4, callus proliferation culture：
The callus successfully induced is seeded on test medium, 5 examination materials of every bottle of inoculation, each processing inoculation 30
Bottle weighs every bottle of callus fresh weight, weighs again after cultivating 45d, and 3 repetitions count callus proliferative conditions, filter out jatamans valeriana rhizome and be cured
Injured tissue is proliferated optimum medium；
S5, culture of rootage：
By+0.5~2.0 mgL of well-grown jatamans valeriana rhizome tissue-cultured seedling access 1/2MS minimal mediums-1IAA is cultivated,
Each 20 bottles of processing inoculation, 3 repetitions, 60 d are 1 period, observe and count the situation of taking root of jatamans valeriana rhizome seedling strong sprout,
Filter out the culture medium of suitable jatamans valeriana rhizome seedling strengthening seedling and rooting culture；
S6, acclimatization and transplants：
Wait for that jatamans valeriana rhizome tissue-cultured seedling grows to 4~5cm, when 3~4cm of root long, transplanting is to heliogreenhouse after 3~5 d of room temperature lower refining seedling
In；Cultivation temperature is 25 ± 2 DEG C, humidity 70%~80%；
Cultivation matrix is turf：Perlite（1：1）, observe growing state, statistics transplanting survival rate.
2. a kind of medicinal plant jatamans valeriana rhizome tissue culture mating system according to claim 1, it is characterised in that：In the S3
Sterile cleaning is to handle 30s to lay-by material using 75% alcohol, aseptic water washing 3~5 times, then with 0.1% HgCl2Fully
10~15min is impregnated, sterile water rinses 4~6 times repeatedly, and fully shaking is by HgCl2It removes, explant body surface is blotted with aseptic filter paper
Face moisture, and be unfolded to place spare.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109220795A (en) * | 2018-10-09 | 2019-01-18 | 云南省农业科学院药用植物研究所 | A kind of jatamans valeriana rhizome tissue culture culture medium and cultural method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109220795A (en) * | 2018-10-09 | 2019-01-18 | 云南省农业科学院药用植物研究所 | A kind of jatamans valeriana rhizome tissue culture culture medium and cultural method |
| CN109220795B (en) * | 2018-10-09 | 2022-01-25 | 云南省农业科学院药用植物研究所 | Tissue culture medium and culture method for valeriana jatamansi jones |
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