CN107047310A - A kind of cultural method of bletilla striata seeds culture seedling - Google Patents
A kind of cultural method of bletilla striata seeds culture seedling Download PDFInfo
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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
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- A01H4/008—Methods for regeneration to complete plants
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Abstract
A kind of cultural method of bletilla striata seeds culture seedling, is related to a kind of cultural method of plant seed seedling, and specially bletilla striata seeds cultivate seedling culture method；Including seed collection, storage；Sterilization, induction seedling, strengthening seedling and rooting, hardening are with transplanting 5 steps；Can a kind of high method of efficiency high, survival rate be provided for bletilla striata tissue cultures by the tissue culture method of the sterile induction seedling of bletilla striata seeds, it is rapid in a short time to obtain the largely strong bletilla striata seedling of stalwartnesses, safety, viability.
Description
Technical field
The present invention relates to a kind of cultural method of plant seed seedling, specially bletilla striata seeds cultivate seedling culture side
Method.
Background technology
The bletilla striata (Bletilla striata(Thunb.) Reichb. f.) belong to perennial herb for the orchid family bletilla striata
Plant, nascent pseudobulb is spheroidal, and growth just forms the block pseudobulb of V-shaped to a certain extent；4~6 pieces of leaf, long and narrow circle
Or lanceolar, long 8~29 centimetres, wide 1.5~4 centimetres, tip is tapering, and base portion, which is received, narrow into sheath and embraces stem；Inflorescence has 3~10
Flower, often not branch or extremely rare branch；Rachis is more or less tortuous in " it " shape；Petal length of a film round shape lanceolar, long by 2~
2.5 centimetre；Hua great, aubergine or pink；4~May of florescence；8~November of fruiting period；The seed of the bletilla striata is superfine small, like powder, shape
There is no endosperm, the embryo of the ateliosis of only several cellularities；Under field conditions (factors), seed germination rate is low.The block of the bletilla striata
Stem is China's traditional Chinese medicine, the effects such as with tonifying lung, detumescence, myogenic, hemostasis, sore, and hindering hemoptysis, metal-inflicted wound for treating lung goes out
The diseases such as blood, canker sores, soup fire burn, rhagadia manus et pedis.
Sprouted using the sterile induction of bletilla striata seeds and produce seedling, then can obtain bletilla seed by being transplanted after strengthening seedling and rooting
Seedling.This breeding method can quickly obtain bletilla striata seedling, can substantially shorten the breeding cycle, and the seedling cultivated is healthy and strong, root system hair
Reach, transplanting survival rate is high.
At present in the bletilla striata tissue-culturing rapid propagation seedling-raising technique reported, it is necessary to selected by explant, protocorm Fiber differentiation,
The production links such as adventitious buds proliferation culture, rooting induction and acclimatization and transplantses can just mass-produce seedling.This tissue culture breeds body
System, middle production link is more, and the production cycle is long, and recruitment cost is high；By protocorm differentiation and adventitious buds proliferation culture, though it can increase
Plus breeding coefficient, but tissue-cultured seedling seedling compared with the seedling that this technology is cultivated is weaker, root system is undeveloped, and transplanting survival rate is low.
The content of the invention
It is an object of the invention to overcome the defect that existing bletilla striata tissue culture technology is present, obtaining one kind can be by white
Splendid achnatherum seed asepsis induces the tissue culture method of seedling, and a kind of high method of efficiency high, survival rate is provided for bletilla striata tissue cultures,
It is rapid in a short time to obtain the strong bletilla striata seedling of a large amount of stalwartnesses, safety, viability.
The purpose of the present invention is achieved through the following technical solutions：
1st, a kind of cultural method of bletilla striata seeds culture seedling, comprises the following steps：
（1）Seed collection, storage：Gather ripe bletilla striata capsule and sealing preserve is standby in 4 DEG C of refrigerator storage；The bletilla striata
The actual storage quantity of capsule is the 120~130% of plan application rate；（Due to the extension with the storage kind time, seed germination rate meeting
It has been declined that, so in storage kind, can increase by 20~30% storage kind amounts, what seed germination rate declined during solution storage kind is made
Into consumption;）
（2）Sterilization：Uncracked bletilla striata capsule is therefrom chosen, is rinsed with flowing water after 30~40min on superclean bench
10~15min is soaked with 75% 20~40s of ethanol postincubation, then with 0.1% mercuric chloride, then with rinsed with sterile water 4~5 times, every time 1
~2min, then blots capsule surface moisture standby with sterile dry filter paper；
（3）Induce seedling：On superclean bench, picking seed is inoculated with the bletilla striata capsule treated with transfer needle from previous step
Onto inducing culture, after light culture to 13~17d, then 60~70d of optical culture, true with more than 2 up to being differentiated to form
Leaf, the seed bud with root；The Medium's PH Value is 5.8 ± 0.2, and cultivation temperature is 24 ± 2 DEG C, and intensity of illumination is
1200lx, light application time 12h/d；According to production needs, to June next year since annual September, monthly induction is sprouted a collection of
Seed, enough seedlings are provided for strengthening seedling and rooting；
(4) strengthening seedling and rooting：Aseptic seedling obtained by previous step is transferred in strengthening seedling and rooting culture medium and carries out culture of rootage, until shape
Into the intact plant with spheroidal pseudobulb；The culture medium pH value is 5.8 ± 0.2, and cultivation temperature is 24 ± 2 DEG C, illumination
Intensity is 1200lx, light application time 12h/d；
(5) hardening is with transplanting：The aseptic bottle of intact plant obtained by previous step is placed in 15~20d of hardening in cool canopy, then in shade
Opened in canopy after bottle cap places 3~5d and take out seedling, cleaned root culture medium, be transplanted at once in sterilized matrix, treat group
After the expansion of training transplanted seedling young leaves and new root are grown, you can routinely manage.
Further：Described（3）Fiber differentiation based component is in step：MS+ benzyladenines 6-BA0.5mg/L+ α-naphthalene
The sucrose of acetic acid NAA0.1mg/L+2.5%.
Further：Described（4）Fiber differentiation based component is in step：MS+ benzyladenines 6-BA0.5mg/L+ α-naphthalene second
The sour sucrose of NAA1.5mg/L+0.5mg/L cycocels+2.5%.
Further：Described（1）The acquisition time of bletilla striata capsule is 9~October in step, when bletilla striata capsule is brown by yellowish turn
When color is ripe.
Further：Described（5）Cool canopy degree of shading is 60~80% in step.
Advantageous effects
The present invention is to make full use of abundant bletilla striata seeds resource, by tissue culture Sterile Culture Methods Used, is existed solving bletilla striata seeds
Under natural conditions germination and growth seedling is directly induced while germination rate low technical problem, by it；Of the invention and existing tissue culture
Technology is compared, and mainly has following three aspect different：One be sprouted by the induction of seed, seedling of directly growing up, reduce tissue culture
The links such as protocorm Fiber differentiation, adventitious buds proliferation culture in production process, shorten the nursery production cycle, save recruitment into
This；Two be, using abundant bletilla striata seeds resource, to be needed to carry out aseptically sowing seeds in batches according to production, be next production
Link provides sufficient introduces a collection；Three be that, by seedling produced by the invention, different from the tissue-cultured seedling that traditional tissue culture is produced, category seed is lured
The seedling of sprouting is led, robust plant, well developed root system, transplant survival are high.
Embodiment
Operating technology of the present invention is further illustrated with reference to example.
Embodiment 1：A kind of cultural method of bletilla striata seeds culture seedling, comprises the following steps：
(1) seed collection, storage：The 9-10 months, when bletilla striata capsule by it is yellowish turn brown it is ripe when, gather ripe bletilla striata capsule
Simultaneously sealing preserve is standby in 4 DEG C of refrigerator storage for fruit；The actual storage quantity of the bletilla striata capsule is the 120% of plan application rate；
(2) sterilization：Take and the seed of 1 month is preserved with embryo age in April, in 4 DEG C of refrigerators, as lay-by material, from standby
With uncracked bletilla striata capsule is chosen in material, 30min is rinsed with flowing water, 75% ethanol postincubation is used on superclean bench
10min is soaked with 0.1% mercuric chloride again after 30s, mercuric chloride solution is outwelled, with rinsed with sterile water 4 times, each 1min uses sterile drying
It is standby that filter paper blots capsule surface moisture；
(3) seedling is induced：On superclean bench, picking seed is inoculated with the bletilla striata capsule treated with transfer needle from previous step
Onto inducing culture, light culture to 13d, then optical culture 60d, until being differentiated to form with more than 2 true leaves, with root
Seed bud, Fiber differentiation based component is preferably：MS+ benzyladenine 6-BA0.5mg/L+ α-naphthylacetic acids NAA0.1mg/L+
2.5% sucrose；Medium's PH Value is 5.6, and cultivation temperature is 22 DEG C, and intensity of illumination is 1200lx, light application time 12h/
d；According to production needs, to June next year since annual September, batch of seeds is sprouted in monthly induction, and foot is provided for strengthening seedling and rooting
Enough seedlings；
(4) strengthening seedling and rooting：Aseptic seedling obtained by previous step is transferred to progress strengthening seedling and rooting culture in strengthening seedling and rooting culture medium, directly
To intact plant of the formation with spheroidal pseudobulb；Strengthening seedling and rooting medium component is preferably：MS+ benzyladenines 6-
BA0.5mg/L+ α-the sucrose of methyl α-naphthyl acetate NAA1.5mg/L+0.5mg/L cycocels+2.5%；Medium's PH Value is 5.6, culture
Temperature is 22 DEG C, and intensity of illumination is 1200lx, light application time 12h/d.
(5) hardening is with transplanting：The aseptic bottle of intact plant obtained by previous step is placed in the cool canopy that degree of shading is 60% and refined
Seedling 15d, then opens bottle cap in cool canopy and places 3d, seedling is taken out, clean root culture medium and be transplanted to sterilized base at once
In matter, appropriateness shade keeps certain humidity, after the expansion of tissue culture transplanted seedling young leaves and new root are grown, you can routinely manage.
Embodiment 2：A kind of cultural method of bletilla striata seeds culture seedling, comprises the following steps：
(1) seed collection, storage：The 9-10 months, when bletilla striata capsule by it is yellowish turn brown it is ripe when, gather ripe bletilla striata capsule
Simultaneously sealing preserve is standby in 4 DEG C of refrigerator storage for fruit；The actual storage quantity of the bletilla striata capsule is the 130% of plan application rate；
(2) sterilization：Take and the seed of 1 month is preserved with embryo age in April, in 4 DEG C of refrigerators, as lay-by material, from standby
With uncracked bletilla striata capsule is chosen in material, 40min is rinsed with flowing water, 75% ethanol postincubation is used on superclean bench
15min is soaked with 0.1% mercuric chloride again after 30s, mercuric chloride solution is outwelled, with rinsed with sterile water 5 times, each 2min uses sterile drying
It is standby that filter paper blots capsule surface moisture；
(3) seedling is induced：On superclean bench, picking seed is inoculated with the bletilla striata capsule treated with transfer needle from previous step
Onto inducing culture, light culture to 17d, then optical culture 70d, until being differentiated to form with more than 2 true leaves, with root
Seed bud, Fiber differentiation based component is preferably：MS+ benzyladenine 6-BA0.5mg/L+ α-naphthylacetic acids NAA0.1mg/L+
2.5% sucrose；Medium's PH Value is 6, and cultivation temperature is 26 DEG C, and intensity of illumination is 1200lx, light application time 12h/d；According to
Production is needed, to June next year since annual September, and batch of seeds is sprouted in monthly induction, and enough kinds are provided for strengthening seedling and rooting
Seedling；
(4) strengthening seedling and rooting：Aseptic seedling obtained by previous step is transferred to progress strengthening seedling and rooting culture in strengthening seedling and rooting culture medium, directly
To intact plant of the formation with spheroidal pseudobulb；Strengthening seedling and rooting medium component is preferably：MS+ benzyladenines 6-
BA0.5mg/L+ α-the sucrose of methyl α-naphthyl acetate NAA1.5mg/L+0.5mg/L cycocels+2.5%；Medium's PH Value is 6, culture temperature
Spend for 26 DEG C, intensity of illumination is 1200lx, light application time 12h/d.
(5) hardening is with transplanting：The aseptic bottle of intact plant obtained by previous step is placed in the cool canopy that degree of shading is 80% and refined
Seedling 20d, then opens bottle cap in cool canopy and places 5d, seedling is taken out, clean root culture medium and be transplanted to sterilized base at once
In matter, appropriateness shade keeps certain humidity, after the expansion of tissue culture transplanted seedling young leaves and new root are grown, you can routinely manage.
Embodiment 3：A kind of cultural method of bletilla striata seeds culture seedling, comprises the following steps：
(1) seed collection, storage：The 9-10 months, when bletilla striata capsule by it is yellowish turn brown it is ripe when, gather ripe bletilla striata capsule
Simultaneously sealing preserve is standby in 4 DEG C of refrigerator storage for fruit；The actual storage quantity of the bletilla striata capsule is the 125% of plan application rate；
(2) sterilization：Take and the seed of 1 month is preserved with embryo age in April, in 4 DEG C of refrigerators, as lay-by material, from standby
With uncracked bletilla striata capsule is chosen in material, 35min is rinsed with flowing water, 75% ethanol postincubation is used on superclean bench
13min is soaked with 0.1% mercuric chloride again after 30s, mercuric chloride solution is outwelled, with rinsed with sterile water 4 times, each 1min uses sterile drying
It is standby that filter paper blots capsule surface moisture；
(3) seedling is induced：On superclean bench, picking seed is inoculated with the bletilla striata capsule treated with transfer needle from previous step
Onto inducing culture, light culture to 15d, then optical culture 65d, until being differentiated to form with more than 2 true leaves, with root
Seed bud, Fiber differentiation based component is preferably：MS+ benzyladenine 6-BA0.5mg/L+ α-naphthylacetic acids NAA0.1mg/L+
2.5% sucrose；Medium's PH Value is 5.8, and cultivation temperature is 24 DEG C, and intensity of illumination is 1200lx, light application time 12h/d；Root
Needed according to production, to June next year since annual September, batch of seeds is sprouted in monthly induction, is provided enough for strengthening seedling and rooting
Seedling；
(4) strengthening seedling and rooting：Aseptic seedling obtained by previous step is transferred to progress strengthening seedling and rooting culture in strengthening seedling and rooting culture medium, directly
To intact plant of the formation with spheroidal pseudobulb；Strengthening seedling and rooting medium component is preferably：MS+ benzyladenines 6-
BA0.5mg/L+ α-the sucrose of methyl α-naphthyl acetate NAA1.5mg/L+0.5mg/L cycocels+2.5%；Medium's PH Value is 5.8, culture
Temperature is 24 DEG C, and intensity of illumination is 1200lx, light application time 12h/d.
(5) hardening is with transplanting：The aseptic bottle of intact plant obtained by previous step is placed in the cool canopy that degree of shading is 70% and refined
Seedling 18d, then opens bottle cap in cool canopy and places 4d, seedling is taken out, clean root culture medium and be transplanted to sterilized base at once
In matter, appropriateness shade keeps certain humidity, after the expansion of tissue culture transplanted seedling young leaves and new root are grown, you can routinely manage.
In order to further reflect the technique effect of the present invention, there is provided tests below data：
（1）Kernel maturing and optimal acquisition phase determine
Closely, the maturity of seed is represented the magnitude relationship in seed germination rate, sprout time etc. and embryo age with embryo age, and embryo age is
To ovary increasing into the time required to fruit since pollination.This technology chooses the bletilla striata to collect the seed of optimal maturity
Embryo age be 3,4,5, the embryo of 6 months, aseptic seeding is in Seed inducement culture medium：MS+ 6-BA0.5mg/L+ NAA0.1mg/L
+ 2.5% sucrose；After pH values is 5.8 ± 0.2,25 DEG C of light culture 15d, it is seen that seed expands sprouting, then with 1200lx
The light application time culture 30-60d of intensity of illumination, 12h/d, investigates the germination rate and sprout time of seed, studies seed maturity
Spend the influence sprouted to seed；
Its sprout time reduces, the bletilla striata seeds sprout time of 3 months up to 60 days, 4~5 months with the increase in embryo age
Bletilla striata sprout time be 30 days or so, the sprout time and 3 months seeds of 6 months embryo age seeds are roughly the same；Bletilla striata seeds
Germination rate and planting percent embryo age be 3 months when germination rate and planting percent it is relatively low, all below 30%, embryo age is at 4~5
Month bletilla striata seeds germination percentage and planting percent be held essentially constant, embryo age had in the bletilla striata seeds germination percentage and planting percent of 6 months
Declined, capsule is easy to crack, seriously polluted and slow-growing when embryo age sterilizes more than 75%, but 6 months；
Therefore, consider, bletilla striata axenic germination most suitable embryo age be 4~5 months, sprout time be 28 days, germination rate and
Planting percent highest, respectively 93% and 92%, growth of seedling is best.Illustrate embryo age of 4~5 months（Namely 9~October, when white
Splendid achnatherum capsule by it is yellowish turn brown it is ripe when）, embryonic development is ripe, is relatively more suitable for the axenic germination and seedling of bletilla striata seeds；
（2）Seed is storeed
Seed vigor refers to the vitality that the potential ability or embryo of germination have, and it is a weight of this technology invention
Want technical parameter.Capsule of the collection with the embryo age of 3~4 months, the dust on surface is washed, at room temperature placement 2 days, then
Part capsule is put in kraft paper bag, is placed under 3 kinds of temperature conditionss and makees storage processing：20 DEG C of refrigerators；4 DEG C of refrigerators；Room temperature bar
Part；Storage time be 1,3,6 months, storage period 1,3 and after 6 months grab sample make germination determine and vitality test；
By experiment, from the point of view of period of storage, when bletilla striata seeds are stored one month, seed vitality during relatively new collection is high, may
Reason is that bletilla striata seeds have after ripening, and when storing 3 months, seed vitality has declined, when storing 6 months, and seed is sprouted
Rate declines by a big margin.Three kinds of banking systems, preferably, after storing one month, seed germination rate is 99% to 4 DEG C of freezer storage effects,
Seed rate of dyeing is 99%, and when storing 6 months, germination rate is more than 70%；Other two kinds of banking systems, seed germination rate and vigor
Decline by a big margin, storage effect is poor；
It follows that 4 DEG C of freezer storages are the best modes preserved the bletilla striata stem of noble dendrobium seed short time；
（3）Sterilization
Saved backup from 4 DEG C of refrigerators are sealed in and uncracked bletilla striata capsule chosen in material, 30~40min is rinsed with flowing water,
With 10~15min is soaked with 0.1% mercuric chloride again after 75% ethanol postincubation 30s on superclean bench, mercuric chloride solution is outwelled, is used
Rinsed with sterile water 4~5 times, 1~2min, blots capsule surface moisture standby with sterile dry filter paper every time；
（4）Seed asepsis induces seedling
On superclean bench, with transfer needle, picking seed is inoculated on inducing culture from sterilized bletilla striata capsule, secretly
Cultivate to 13~17 d, then 60~70d of optical culture, until being differentiated to form with more than 2 true leaves, the seed seedlings with root；
Fiber differentiation based component is preferably：MS+ benzyladenines 6-BA0.5mg/L+ α-methyl α-naphthyl acetate NAA0.1mg/L
+ 2.5% sucrose；Culture medium pH value is 5.8 ± 0.2, and cultivation temperature is 24+2 DEG C, and intensity of illumination is 1200lx, illumination
Time 12h/d；
According to production needs, to June next year since September, batch of seeds is sprouted in monthly sterile induction, and foot is provided for strengthening seedling and rooting
Enough seedlings；
（5）Strengthening seedling and rooting
Aseptic seedling obtained by previous step is transferred to progress strengthening seedling and rooting culture in strengthening seedling and rooting culture medium, ball is carried until being formed
The intact plant of shape pseudobulb；
Strengthening seedling and rooting medium component is preferably：MS+ benzyladenines 6-BA0.5mg/L+ α-methyl α-naphthyl acetate NAA1..5mg
/ L+0.5mg/the sucrose of L cycocels+2.5%；Culture medium pH value is 5.8 ± 0.2, and cultivation temperature is 24+2 DEG C, light
It is 1200lx, light application time 12h/d according to intensity；
（6）Hardening is with transplanting
By the aseptic bottle of above-mentioned acquisition intact plant, degree of shading is placed in up to 15~20d of hardening in 60~80% cool canopies, then in shade
Bottle cap is opened in canopy and places 3~5d, seedling is taken out, root culture medium is cleaned and is transplanted at once in sterilized matrix, appropriateness hides
Shade, keeps certain humidity, after the expansion of tissue culture transplanted seedling young leaves and new root are grown, you can routinely manage.
Without departing from the present invention, various conversion and equivalent replacement can also be carried out to invention, therefore, this
Patent of invention is not limited to disclosed specific implementation process, and should include the whole that falls within the scope of the appended claims
Embodiment.
Claims (5)
1. a kind of cultural method of bletilla striata seeds culture seedling, it is characterised in that comprise the following steps：
（1）Seed collection, storage：Gather ripe bletilla striata capsule and sealing preserve is standby in 4 DEG C of refrigerator storage；The bletilla striata
The actual storage quantity of capsule is the 120~130% of plan application rate；
（2）Sterilization：Uncracked bletilla striata capsule is therefrom chosen, is rinsed with flowing water after 30~40min on superclean bench
10~15min is soaked with 75% 20~40s of ethanol postincubation, then with 0.1% mercuric chloride, then with rinsed with sterile water 4~5 times, every time 1
~2min, then blots capsule surface moisture standby with sterile dry filter paper；
（3）Induce seedling：On superclean bench, picking seed is inoculated with the bletilla striata capsule treated with transfer needle from previous step
Onto inducing culture, after light culture to 13~17d, then 60~70d of optical culture, true with more than 2 up to being differentiated to form
Leaf, the seed bud with root；The Medium's PH Value is 5.8 ± 0.2, and cultivation temperature is 24 ± 2 DEG C, and intensity of illumination is
1200lx, light application time 12h/d；According to production needs, to June next year since annual September, monthly induction is sprouted a collection of
Seed, enough seedlings are provided for strengthening seedling and rooting；
(4) strengthening seedling and rooting：Aseptic seedling obtained by previous step is transferred in strengthening seedling and rooting culture medium and carries out culture of rootage, until shape
Into the intact plant with spheroidal pseudobulb；The culture medium pH value is 5.8 ± 0.2, and cultivation temperature is 24 ± 2 DEG C, illumination
Intensity is 1200lx, light application time 12h/d；
(5) hardening is with transplanting：The aseptic bottle of intact plant obtained by previous step is placed in 15~20d of hardening in cool canopy, then in shade
Opened in canopy after bottle cap places 3~5d and take out seedling, cleaned root culture medium, be transplanted at once in sterilized matrix, treat group
After the expansion of training transplanted seedling young leaves and new root are grown, you can routinely manage.
2. the cultural method of a kind of bletilla striata seeds culture seedling according to claim 1, it is characterised in that described（3）
Fiber differentiation based component is in step：The MS+ benzyladenines 6-BA0.5mg/L+ α-sugarcanes of methyl α-naphthyl acetate NAA0.1mg/L+2.5%
Sugar.
3. a kind of cultural method of bletilla striata seeds culture seedling according to claims 1 or 2 any one, its feature exists
In described（4）Fiber differentiation based component is in step：MS+ benzyladenine 6-BA0.5mg/L+ α-naphthylacetic acids NAA1.5mg/L
The sucrose of+0.5mg/L cycocels+2.5%.
4. the cultural method of a kind of bletilla striata seeds culture seedling according to claim 3, it is characterised in that described（1）
The acquisition time of bletilla striata capsule is 9~October in step, when bletilla striata capsule by it is yellowish turn brown it is ripe when.
5. the cultural method of a kind of bletilla striata seeds culture seedling according to claim 4, it is characterised in that described（5）
Cool canopy degree of shading is 60~80% in step.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108308031A (en) * | 2018-04-16 | 2018-07-24 | 安徽东方金桥农林科技股份有限公司 | A kind of pale reddish brown trident bletilla seed tissue cultural method |
| CN109076923A (en) * | 2018-08-28 | 2018-12-25 | 河南云帮农业科技有限公司 | A kind of bletilla striata aseptic seeding special culture media |
| CN109452171A (en) * | 2018-11-26 | 2019-03-12 | 丽江海贝瑞生物科技有限公司 | A kind of tissue culture method of the sterile induction plant regeneration of pale reddish brown trident bletilla striata seeds |
| CN110089429A (en) * | 2019-04-25 | 2019-08-06 | 浙江省农业科学院 | A method of quickly breeding bletilla seedling using method for tissue culture |
| CN111903530A (en) * | 2020-09-23 | 2020-11-10 | 宣威市福康生物科技有限公司 | Tissue culture seedling method for bletilla striata |
| CN112042537A (en) * | 2020-09-09 | 2020-12-08 | 三峡大学 | Method for establishing bletilla striata plant regeneration system |
| CN116649216A (en) * | 2023-06-25 | 2023-08-29 | 江苏护理职业学院 | Pure bletilla striata three-fork tissue culture domesticated seedling raising method |
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| CN109452171A (en) * | 2018-11-26 | 2019-03-12 | 丽江海贝瑞生物科技有限公司 | A kind of tissue culture method of the sterile induction plant regeneration of pale reddish brown trident bletilla striata seeds |
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| CN112042537A (en) * | 2020-09-09 | 2020-12-08 | 三峡大学 | Method for establishing bletilla striata plant regeneration system |
| CN112042537B (en) * | 2020-09-09 | 2022-04-08 | 三峡大学 | Method for establishing bletilla striata plant regeneration system |
| CN111903530A (en) * | 2020-09-23 | 2020-11-10 | 宣威市福康生物科技有限公司 | Tissue culture seedling method for bletilla striata |
| CN116649216A (en) * | 2023-06-25 | 2023-08-29 | 江苏护理职业学院 | Pure bletilla striata three-fork tissue culture domesticated seedling raising method |
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