CN106258976B - A kind of tissue culturing fast seedling-cultivating method of mustard type rape - Google Patents
A kind of tissue culturing fast seedling-cultivating method of mustard type rape Download PDFInfo
- Publication number
- CN106258976B CN106258976B CN201610714886.5A CN201610714886A CN106258976B CN 106258976 B CN106258976 B CN 106258976B CN 201610714886 A CN201610714886 A CN 201610714886A CN 106258976 B CN106258976 B CN 106258976B
- Authority
- CN
- China
- Prior art keywords
- culture
- callus
- explant
- adventitious bud
- seedling
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
-
- 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
- A01H4/001—Culture apparatus for tissue culture
-
- 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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The present invention relates to a kind of tissue culturing fast seedling-cultivating methods of mustard type rape, belong to Plant Tissue Breeding quick breeding technology field.Including following the step of sequentially carrying out:（1）The acquisition of aseptic explant;（2）Explant dedifferentiation is induced to form callus;（3）Evoked callus is differentiated to form adventitious bud again;（4）The squamous subculture of adventitious bud；（5）Culture of rootage；（6）Seedling acclimatization and transplants.Using the method for the invention, the dedifferentiation frequency of explant is more than 99%, and for the differentiation rate again of callus more than 35%, pollution rate is few, and the healthy and strong tissue-cultured seedling of the qualification that all can be used for plantation of acquisition, transplanting survival rate is more than 98%.
Description
Technical field
The present invention relates to a kind of methods of mustard type rape tissue cultures and fast seedling growing, and it is quick to belong to Plant Tissue Breeding
Reproduction technique field.
Technical background
Rape is one of the important oil crops in the world, including turnip type rape (genome AA), mustard type rape (gene
Group AABB) and three cultigens of cabbage type rape (genome AACC).Plant tissue culture technique had more than 100 years so far
History, and technology reaches its maturity and perfect, played an important role in the genetic improvement of crops.With working as molecular biosciences
Development, the tissue culture technique of rape have been more and more widely used in production and research work.For example, rape
In genetic breeding haploid breeding and fast breeding are carried out by tissue cultures；Gene function is tested in research and is obtained by tissue cultures
Obtain genetic transformation positive plant etc..In Oil Rape Tissue incubation, stem, rachis, Cotyledons with petiole, hypocotyl, blade and small
Spore etc. can serve as explant and carry out in-vitro inducing acquisition regeneration plant.But it is found in specific implementation process
It is very big in different genotype kind, the differentiation efficiency difference of different types of Leaf Stems from Brassica juncea, obtain regrowth technical system
Poor repeatability.At present in the tissue culture procedures of mustard type rape, due to different genotype kind, explant type and group
The influence of various hormone ratios in cultivating system is knitted, the efficiency variance for obtaining regrowth is still very big.There are one ripe
Reliable mustard type rape tissue culture technique system obtains a large amount of mustard type rape plant regeneration seedling.
Invention content
The purpose of the present invention is to provide a kind of tissue culturing fast seedling-cultivating methods of mustard type rape, now raw to solve
The above problem present in production, method provided by the present invention is easy to operate, can obtain the excellent plant of a large amount of characters, input
Few, seedling is quick, plants robust seedling, and research platform is provided for the deep genetic transformation for carrying out rape and character improvement.
To achieve these goals, the present invention takes following technical measures：
A kind of tissue culturing fast seedling-cultivating method of mustard type rape, including：
(1) acquisition of aseptic explant：Mustard type rape is selected to be sterilized on superclean bench into real full seed,
Seed after sterilizing is seeded into seed germination medium, the hypocotyl of elongation is then cut into 7mm- under sterile conditions
The segment explant of 12mm；；
The seed germination medium includes：MS culture mediums+6.0~8.0g/L of agar powder, pH value are 5.8~6.0；It broadcasts
After kind, light culture.
Preferably, the sterilizing of seed is with after 75% alcohol immersion treatment 3min, is put into sterile water and rinses 1min, then
It is put into after impregnating 10min in 0.1% mercuric chloride, aseptic water washing 6-7 times；
(2) induction explant dedifferentiation forms callus：Under sterile conditions, the explant cut is put into callus
In inducing culture, induction explant forms callus；
The formula of calli induction media includes：MS culture mediums+16~20g/L+2,4- of sucrose 28~32g/L+ mannitol
D0.8~1.2mg/L+Kinetin 0.25~0.35mg/L+ agar powder 6g/L, pH value are 5.8~6.0.
(3) evoked callus is differentiated to form indefinite bud point again：It under sterile conditions, will be obtained in step (2)
Callus is inserted into indefinite bud point inducing culture, and induction shape callus forms indefinite bud point；
The formula of indefinite bud point inducing culture includes：MS culture mediums+28~32g/L+TDZ1.2 of sucrose~1.8g/L+
NAA0.08~1.2g/L+ silver nitrate 4.8~5.2mg/L+, 7.0~7.4g/L of agar powder, pH value are 5.8~6.0；
(4) grown cultures of adventitious bud：Aseptically, the callus with indefinite bud point that will be obtained in step (3)
Tissue is transferred in adventitious bud growth medium, carries out the grown cultures of adventitious bud；
The formula of adventitious bud growth medium includes：MS culture mediums+0.23~0.27g/ of glucose 8.0~12g/L+ xyloses
0.5~0.7g/L+ of L+MES, 0.08~0.12mg/L+ of trans- 1.8~2.2mg/L+IAA of-Zeatin agar powders 5.8~
6.2g/L, pH value are 5.8~6.0；
(5) culture of rootage：Aseptically, the adventitious bud that step (4) obtains is cut into single plant, is inoculated into training of taking root
It supports and carries out culture of rootage on base；
The formula of root media includes：MS culture mediums+sucrose 8~12g/L+, 8~12g/L of agar powder, pH value for 5.8~
6.0；
(6) Multiplying culture：The adventitious bud being inoculated on root media grows up to the complete plant with multiple collateral generation points
Strain after the plant is cut into the 2-3 sections of stem sections with collateral generation point, is inoculated into increment culture medium and carries out increment culture, expand
Numerous plant；Subculture is primary within two weeks.
The formula of the proliferated culture medium is identical with root media；
Approach described above, the cultivation temperature of step (2)-(6) is 20~24 DEG C, illuminance 2000-2500lx, illumination 15
~18 hours/day.
(7) seedling acclimatization and transplants：The seedling taken root obtained in step (6) is subjected to hardening, then routinely technology
It is transplanted in crop field or greenhouse.
Published all mustard type rape strains can carry out nursery in aforementioned manners.
Compared with prior art, beneficial effects of the present invention：
(1) present invention can be obtained big by the use of the hypocotyl of rape as explant in the case where using less seed
The elite plant of amount.
(2) present invention carrys out the formation of evoking adventive bud point using adventitious bud induction culture base, substantially increases callus
The efficiency broken up again, the differentiation rate of callus reach more than 35%.
(3) present invention has induced the indefinite bud point through formation to be grown to serve as normally using adventitious bud growth medium to cultivate
Adventitious bud, which greatly enhances the growth efficiencies of the adventitious bud containing growing point.
(4) the characteristics of present invention is easy to operate, has less investment, and output is high.Breeding cycle is short, and the young plant of acquisition is strengthened, root
It is more, transplanting survival rate is up to more than 98%.
The hypocotyl segment of (5) long 1cm passes through the culture of 2 weeks, can manage it into one piece of area for 0.3cm2Callus group
It knits, by the culture of 2 weeks, 1-3 indefinite bud points can be formed, then through the culture of 3-4 weeks, each adventitious bud clump can obtain one
The adventitious bud that can be transferred to above in root media.Adventitious bud is cultivated after two weeks in root media, can obtain 2-3 sections
Stem section with collateral generation point is inoculated into root media and carries out culture of rootage.By the culture of rootage of 4-5 weeks, can obtain
The 2-3 seedling taken root is obtained, proliferation times are 2-3 times, and an adventitious bud can breed tens million of or more seedling within 1 year.With one
A bud 14d subcultures are primary, and each proliferation times are calculated for 3.
Description of the drawings
Fig. 1 is that explant dedifferentiation is callus.
Fig. 2 is divided into indefinite bud point for evoked callus (arrow show indefinite bud point).
Fig. 3 is the upgrowth situation that indefinite bud point after two weeks is cultivated on adventitious bud growth medium.
Seedling upgrowth situation in Fig. 4 root medias.
Fig. 5 observes situation of taking root for bottom of bottle.
Specific embodiment
Technical solution of the present invention is the ordinary skill in the art if not otherwise specified.The reagent or material,
If not otherwise specified, commercial channel is derived from.
Embodiment 1：
The tissue culturing fast seedling-cultivating method of mustard type rape strain J248 (Xu et al.2014)：
(1) acquisition of explant：Mustard type rape strain J248 is selected to be used on superclean bench into real full seed
After 75% alcohol impregnates 3min, it is put into sterile water and rinses 1min.It is then placed in after impregnating 10min in 0.1% mercuric chloride, sterile water
It rinses 6-7 times.Seed after sterilizing is seeded into seed germination medium, after planting, light culture 7d.Then by the lower embryo of elongation
Axis is cut into the segment explant of 1cm or so under sterile conditions.
The seed germination medium is：MS culture medium+7g/L agar powders, pH value 5.9.
(2) induction explant dedifferentiation forms callus：Under sterile conditions, the explant cut is put into callus
In inducing culture, after 12d, more than 99% explant initially forms callus.
The calli induction media includes：MS culture medium+30g/L sucrose+18g/L mannitol+1mg/L 2,4-D+
0.3mg/L Kinetin+6g/L agar powders, pH value 5.9.
(3) evoked callus is differentiated to form indefinite bud point again：The explant cut is put into calli induction media 14 days
Afterwards, under sterile conditions, the callus of acquisition is inserted into indefinite bud point inducing culture, after 10 days, 44% callus
Tissue initially forms indefinite bud point.
The indefinite bud point inducing culture includes：MS culture medium+30g/L sucrose+1.5g/L TDZ+0.1g/L NAA
+ 5.0mg/L silver nitrate+7g/L agar powders, pH value 5.9.
(4) grown cultures of adventitious bud：Aseptically, the callus with indefinite bud point that will be obtained in step (3)
Tissue is transferred in adventitious bud growth medium, carries out grown cultures.It does not grow the explant of indefinite bud point also, then continues to put
Fiber differentiation is carried out into indefinite bud point inducing culture, after indefinite bud point is grown, is transferred in adventitious bud growth medium.
Adventitious bud growth medium includes：MS culture medium+10g/L glucose+0.25g/L xylose+0.6g/L MES+
Trans--Zeatin+0.1mg/L IAA+6g/L the agar powders of 2.0mg/L, pH value 5.9.
(5) culture of rootage：Aseptically, the adventitious bud obtained in step (4) is cut into single plant, is inoculated into and takes root
The culture of rootage of 30 days is carried out on culture medium.Rooting rate is 100%.
The root media includes：MS culture medium+10g/L sucrose+10g/L agar powders, pH value 5.9.
(6) Multiplying culture：The adventitious bud being inoculated on root media grows up to the complete of multiple collateral generation points
Plant after the plant to be cut into the stem section with collateral generation point, is inoculated into increment culture medium and carries out increment culture, expand numerous plant
Strain；Subculture is primary within two weeks.
The formula of the proliferated culture medium is identical with root media；
Approach described above, the cultivation temperature of step (2)-(6) is 22 DEG C, illuminance 2200lx, 16 hour/day of illumination.
(7) seedling acclimatization and transplants：Taken root 50 seedlings obtained in step (6) are subjected to hardening, then routinely
Technology is transplanted in greenhouse, and the survival rate of small transplantation of seedlings is 98%.In the present embodiment, average each intact plant it is cleavable into
The 2-3 stem sections for carrying collateral generation point.
Embodiment 2：
The tissue culturing fast seedling-cultivating method of mustard type rape strain J163-4 (Xu et al.2014)：
(1) acquisition of explant：Mustard type rape is selected into real full seed, with 75% alcohol on superclean bench
After cleaning 3min, it is put into sterile water and rinses 1min.It is then placed in after cleaning 10min in 0.1% mercuric chloride, aseptic water washing 6-7
It is secondary.Seed after sterilizing is seeded into seed germination medium, after planting, light culture 7d.Then by the hypocotyl of elongation in nothing
The segment explant of 1-2cm is cut under conditions of bacterium.
The seed germination medium includes：MS culture medium+7g/L agar powders, pH value 5.9.
(2) induction explant dedifferentiation forms callus：Under sterile conditions, the explant cut is put into callus
In inducing culture, after 12d, more than 99% explant initially forms callus.
The calli induction media includes：MS culture medium+30g/L sucrose+18g/L mannitol+1.0mg/L 2,4-D
+ 0.3mg/L Kinetin+6g/L agar powders, pH value 5.9.
(3) evoked callus is differentiated to form indefinite bud point again：The explant cut is put into calli induction media 14 days
Afterwards, under sterile conditions, the callus of acquisition is inserted into indefinite bud point inducing culture, after 10 days, 36% callus
Tissue initially forms indefinite bud point.
The indefinite bud point inducing culture includes：MS culture medium+30g/L sucrose+1.5g/L TDZ+0.1g/L NAA
+ 5.0mg/L silver nitrate+7.0g/L agar powders, pH value 5.9.
(4) grown cultures of adventitious bud：Aseptically, the callus with indefinite bud point that will be obtained in step (3)
Tissue is transferred in adventitious bud growth medium, carries out grown cultures.It does not grow the explant of indefinite bud point also, then continues to put
Fiber differentiation is carried out into indefinite bud point inducing culture, after indefinite bud point is grown, is transferred in adventitious bud growth medium.
Adventitious bud growth medium includes：MS culture medium+10g/L glucose+0.25g/L xylose+0.6g/L MES+
Trans--Zeatin+0.1mg/L IAA+6g/L the agar powders of 2.0mg/L, pH value 5.9.
(5) culture of rootage：Aseptically, the adventitious bud obtained in step (5) is cut into single plant, is inoculated into and takes root
The culture of rootage of 32 days is carried out on culture medium.Rooting rate is 100%.
The root media includes：MS culture medium+10g/L sucrose+10g/L agar powders, pH value 5.9.
(6) Multiplying culture：The adventitious bud being inoculated on root media grows up to the complete plant with multiple collateral generation points
Strain after the plant is cut into the 2-3 sections of stem sections with collateral generation point, is inoculated into increment culture medium and carries out increment culture, expand
Numerous plant；Subculture is primary within two weeks.
The formula of the proliferated culture medium is identical with root media；
Approach described above, the cultivation temperature of step (2)-(6) is 22 DEG C, illuminance 2200lx, 16 hour/day of illumination.
(7) seedling acclimatization and transplants：Taken root 124 seedlings obtained in step (6) are subjected to hardening, then routinely
Technology is transplanted in greenhouse, the survival rate 99% of small transplantation of seedlings.
Claims (3)
1. a kind of tissue culturing fast seedling-cultivating method of mustard type rape, including：
(1) acquisition of aseptic explant：Mustard type rape is selected to sterilize, will go out on superclean bench into real full seed
Seed after bacterium is seeded into seed germination medium；After planting, light culture；Then by the hypocotyl of elongation under sterile conditions
It is cut into the segment explant of 7mm-12mm；
The seed germination medium is：MS culture mediums+6.0~8.0g/L of agar powder, pH value are 5.8~6.0；
(2) induction explant dedifferentiation forms callus：Under sterile conditions, the explant cut is put into callus induction
In culture medium, induction explant forms callus；
The formula of calli induction media is：MS culture mediums+16~20g/L+2,4-D of sucrose 28~32g/L+ mannitol 0.8~
1.2mg/L+Kinetin 0.25~0.35mg/L+ agar powder 6g/L, pH value are 5.8~6.0；
(3) evoked callus is differentiated to form indefinite bud point again：Under sterile conditions, by the callus obtained in step (2)
Tissue is inserted into indefinite bud point inducing culture, and evoked callus forms indefinite bud point；
The formula of indefinite bud point inducing culture is：MS culture mediums+28~32g/L+TDZ1.2 of sucrose~1.8g/L+NAA 0.08
~1.2g/L+ silver nitrate 4.8~5.2mg/L+, 7.0~7.4g/L of agar powder, pH value are 5.8~6.0；
(4) grown cultures of adventitious bud：Aseptically, the callus with indefinite bud point that will be obtained in step (3)
It is transferred in adventitious bud growth medium, carries out the grown cultures of adventitious bud；
The formula of adventitious bud growth medium is：MS culture mediums+0.23~0.27g/L+MES of glucose 8.0~12g/L+ xyloses
0.5~0.7g/L+ trans- 1.8~2.2mg/L+IAA of-Zeatin 0.08~0.12mg/L+ agar powders 5.8~6.2g/L, pH
Be worth is 5.8~6.0；
(5) culture of rootage：Aseptically, the adventitious bud that step (4) obtains is cut into single plant, is inoculated into root media
Upper carry out culture of rootage；
The formula of root media is：MS culture mediums+sucrose 8~12g/L+, 8~12g/L of agar powder, pH value are 5.8~6.0；
(6) Multiplying culture：The adventitious bud being inoculated on root media grows up to the intact plant with multiple collateral generation points, will
After the plant is cut into the 2-3 sections of stem sections with collateral generation point, it is inoculated into proliferated culture medium and carries out Multiplying culture, expand numerous plant
Strain；Subculture is primary within two weeks；
The formula of the proliferated culture medium is identical with root media；
(7) seedling acclimatization and transplants：The seedling taken root obtained in step (6) is subjected to hardening, then routinely technology is transplanted
To in crop field or greenhouse.
2. according to the method described in claim 1, the cultivation temperature of step (2)-(6) be 20~24 DEG C, illuminance 2000-
2500lx, 15~18 hour/day of illumination.
3. according to the method described in claim 1, the sterilizing is to be put into 75% alcohol by after seed immersion treatment 3min
1min is rinsed in sterile water, is then placed in after impregnating 10min in 0.1% mercuric chloride, aseptic water washing 6-7 times.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610714886.5A CN106258976B (en) | 2016-08-24 | 2016-08-24 | A kind of tissue culturing fast seedling-cultivating method of mustard type rape |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610714886.5A CN106258976B (en) | 2016-08-24 | 2016-08-24 | A kind of tissue culturing fast seedling-cultivating method of mustard type rape |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106258976A CN106258976A (en) | 2017-01-04 |
| CN106258976B true CN106258976B (en) | 2018-07-10 |
Family
ID=57615058
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610714886.5A Expired - Fee Related CN106258976B (en) | 2016-08-24 | 2016-08-24 | A kind of tissue culturing fast seedling-cultivating method of mustard type rape |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106258976B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107873516A (en) * | 2017-11-14 | 2018-04-06 | 广西壮族自治区药用植物园 | The breeding method of yncaria stem with hooks |
| CN107853178B (en) * | 2017-11-22 | 2020-07-28 | 中国热带农业科学院南亚热带作物研究所 | Method for inducing embryogenic callus |
| CN110845266A (en) * | 2019-11-22 | 2020-02-28 | 顾霆 | Embryonic germ cell dedifferentiation culture medium and preparation method thereof |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8796508B2 (en) * | 2005-11-10 | 2014-08-05 | Pioneer Hi-Bred International, Inc. | Microprojectile bombardment transformation of Brassica |
| CN102388800B (en) * | 2011-07-26 | 2013-04-17 | 南京农业大学 | Light source control method for tissue culture of brassica napus |
| CN103421840A (en) * | 2013-08-01 | 2013-12-04 | 华中农业大学 | Method for improving resistance of rape to Lepidoptera pests by transgenic technology |
| CN103773798B (en) * | 2014-01-08 | 2015-12-30 | 华中农业大学 | PLD ε gene is increasing the application in crop nitrogen nutrition efficiency utilization and seed production |
| CN103773784B (en) * | 2014-01-08 | 2016-02-17 | 华中农业大学 | PLD α 1 gene is increasing the application in crop drought resistance and seed production |
| CN103966258B (en) * | 2014-04-29 | 2016-05-18 | 浙江大学 | A kind of agriculture bacillus mediated cabbage type rape genetic transforming method |
| CN104774850A (en) * | 2015-04-03 | 2015-07-15 | 华中农业大学 | BnNRT1-3 gene overexpression for improving rape nitrogen utilization rate |
-
2016
- 2016-08-24 CN CN201610714886.5A patent/CN106258976B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN106258976A (en) | 2017-01-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101611697B (en) | Virus removal and rapid propagation technology of sweet potato variety 'Shangshu 19' | |
| CN106942051B (en) | A kind of culture medium and propagation method of the tissue-culturing quick-propagation of alum root blade | |
| CN108522277B (en) | Tissue culture seedling raising method for semi-lignified stem segments with buds of 'Hongyang' kiwi fruits | |
| CN101401549B (en) | Once-seedling forming quick propagating method for sugarcane tissue culture | |
| CN107155898A (en) | A kind of dendrobium candidum carries out expanding numerous method using stem section thin slice | |
| CN104521756A (en) | Method for producing trichosanthes tissue culture seedlings | |
| CN102047842A (en) | Method for directly regenerating plants by adopting citrullus lanatus cotyledon nodes | |
| CN104719158A (en) | Method for rapidly establishing medium-sized Chinese pennisetum herb tissue culture regeneration system by taking seeds as explants | |
| CN101715730B (en) | Tissue culture rapid propagation method for sugarcane new Taisugar No.10 | |
| CN106258976B (en) | A kind of tissue culturing fast seedling-cultivating method of mustard type rape | |
| CN106538382B (en) | Method for establishing efficient eremochloa ophiuroides regeneration system by taking young ears as explants | |
| CN103070078A (en) | Rapid propagation method for performing tissue culture by using taro stem tip | |
| CN104938335B (en) | The method that regeneration plant is obtained using oil tea hypocotyls | |
| CN117243119B (en) | Method for rapidly obtaining tetraploid cowpea | |
| CN103583357A (en) | Method for sterile seeding of lithops and establishing regeneration system | |
| CN109452170A (en) | A kind of method of sword-like iris seed root evoked callus culture | |
| CN105613288A (en) | Construction method of rapid Euonymus japonicus L.f. aureo-marginatus Rehd propagation system | |
| CN103155869A (en) | Sweet cherry rootstock Colt tissue culture method | |
| CN102668982B (en) | Tissue culture method of mature wheat embryo | |
| CN105766636B (en) | A kind of peony tissue culture regeneration method | |
| CN115633640B (en) | Method for regenerating adventitious buds of canna test-tube plantlet leaf sheath and application thereof | |
| CN113973658B (en) | Efficient genetic transformation and plant regeneration method for capsicum | |
| CN105284617A (en) | Efficient in-vitro propagation method for inducing root segment of kok-saghyz into seedling by one step | |
| CN104285816A (en) | Rapid propagation method for xanthoceras sorbifolia bunge tissue during culturing | |
| CN109349108A (en) | A kind of sweet tea buckwheat somatic embryo occurs and plant regeneration method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20180710 Termination date: 20190824 |