CN105432465A - Method for improving embryonic callus induction rate and embryo transfer rate of pinus koraiensis - Google Patents
Method for improving embryonic callus induction rate and embryo transfer rate of pinus koraiensis Download PDFInfo
- Publication number
- CN105432465A CN105432465A CN201510771278.3A CN201510771278A CN105432465A CN 105432465 A CN105432465 A CN 105432465A CN 201510771278 A CN201510771278 A CN 201510771278A CN 105432465 A CN105432465 A CN 105432465A
- Authority
- CN
- China
- Prior art keywords
- embryo
- medium
- induction
- callus
- embryonic
- 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.)
- Granted
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
-
- 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
- A01H4/008—Methods for regeneration to complete plants
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 invention relates to a method for improving embryonic callus induction rate and embryo transfer rate of pinus koraiensis, and mainly solves the problems that the embryonic callus induction rate and the embryo transfer rate are low when the pinus koraiensis is subjected to somatic embryogenesis. The method comprises the following steps: firstly, sterilizing materials; secondly, inducing embryonic calli; thirdly, performing subculture and maintenance as well as proliferation on the calli; fourthly, inducing proembryos; fifthly, enabling the somatic embryos to develop and mature, and regenerating plants. According to the method, both the embryonic callus induction rate and the embryo transfer rate are improved when the pinus koraiensis is subjected to the somatic embryogenesis through reasonable proportion and suitable culture conditions of various components in a medium; the induction rate of the embryonic calli can reach 33.3 percent; the addition of inositol reduces the browning degree of the embryonic calli, and helps with the transformation of globular embryos; the embryo transfer rate is 6 to 8 times that of pinus koraiensis without adding the inositol. The method is applied to the forest field.
Description
Technical field
The present invention relates to a kind of method improving Korean pine frequency of embryonic callus induction and turn embryo rate.
Background technology
Korean pine (Pinuskoraiensis) is the extremely important high quality timber seeds in the Northeast of China, is also the huge nut nonwood forest trees of development prospect.Somatic embryo is the basis of Fast-propagation and artificial seed development, significant to genetic improvement.Carry out Korean pine somatic embryo to occur and indefinite bud is studied, both can organize training system for its biological study provide effective, the asexual propagation of elite germplasm material can be directly used in again.But carry out now Korean pine somatic cell when occurring, frequency of embryonic callus induction and to turn embryo rate all lower, seriously constrain the research of the somatic embryo generation aspect of Korean pine.
Summary of the invention
The present invention will solve existing Korean pine when carrying out somatic embryo and occurring, frequency of embryonic callus induction and turn the low problem of embryo rate, provides a kind of method improving Korean pine frequency of embryonic callus induction and turn embryo rate.
A kind of Korean pine frequency of embryonic callus induction that improves of the present invention carries out according to the following steps with the method turning embryo rate:
One, the sterilizing of material
By be in the cleavage polyembryony phase cones of Pinus koraiensis sterilizing after, peel off seed coat, again sterilizing, obtain the zygotic embryo after sterilizing;
Two, embryo callus subculture induction
Zygotic embryo after sterilizing step one obtained is inoculated in medium, under the condition of 23 ~ 25 DEG C, light culture 58 ~ 65d, obtain embryo callus, wherein medium is DCR medium+agar 6.5g/L+L-glutamine 500mg/L+ sucrose 35g/L+NAA2mg/L+6-BA1.5mg/L+ acid hydrolyzed casein 0.8g/L; Medium pH=5.8;
Three, the subculture of callus keeps and propagation
Embryo callus step 2 obtained carries out squamous subculture, obtains the callus after breeding;
Four, the induction of proembryo
Callus after propagation is inoculated into the induction carrying out proembryo in medium, and under the condition of 23 ~ 25 DEG C, light culture 7 ~ 15d, obtains the callus with globular embryo; Wherein medium is DCR medium+acid hydrolyzed casein 500mg/L+L-glutamine 500mg/+ sucrose 20g/L+ agar 6.5g/L+ active carbon 2g/L+ inositol 4g/L; PH=5.8;
Five, the growth of somatic embryo and plant regeneration
Callus with globular embryo is inoculated in medium and carries out body embryonic development, complete and turn embryo process, obtain cotyledonary embryos; Maturation induction is carried out to the cotyledonary embryos obtained simultaneously, obtains mature embryo, then carry out sprouting, plant regeneration and transplanting seedling.
The present invention passes through the rational proportion of each component in medium and suitable condition of culture, make Korean pine when carrying out somatic embryo and occurring, frequency of embryonic callus induction and turn embryo rate and be obtained for raising, frequency of embryonic callus induction can reach 33.3%, the present invention finds that inositol plays an important role to the induction of globular embryo and later stage body embryonic development and maturation, by adding inositol, embryo callus subculture brownization degree is reduced, and more contribute to the conversion of globular embryo, turning embryo rate is 6-8 times that does not add inositol.
Accompanying drawing explanation
Fig. 1 is the photo of embryo callus in embodiment one;
Fig. 2 is the photo of early stage proembryos in embodiment one;
Fig. 3 is the photo of globular embryo in embodiment one;
Fig. 4 and Fig. 5 is the photo of somatic embryo development in embodiment one.
Embodiment
Embodiment one: a kind of Korean pine frequency of embryonic callus induction that improves of present embodiment carries out according to the following steps with the method turning embryo rate:
One, the sterilizing of material
By be in the cleavage polyembryony phase cones of Pinus koraiensis sterilizing after, peel off seed coat, again sterilizing, obtain the zygotic embryo after sterilizing;
Two, embryo callus subculture induction
Zygotic embryo after sterilizing step one obtained is inoculated in medium, under the condition of 23 ~ 25 DEG C, light culture 58 ~ 65d, obtain embryo callus, wherein medium is DCR medium+agar 6.5g/L+L-glutamine 500mg/L+ sucrose 35g/L+NAA2mg/L+6-BA1.5mg/L+ acid hydrolyzed casein 0.8g/L; Medium pH=5.8;
Three, the subculture of callus keeps and propagation
Embryo callus step 2 obtained carries out squamous subculture, obtains the callus after breeding;
Four, the induction of proembryo
Callus after propagation is inoculated into the induction carrying out proembryo in medium, and under the condition of 23 ~ 25 DEG C, light culture 7 ~ 15d, obtains the callus with globular embryo; Wherein medium is DCR medium+acid hydrolyzed casein 500mg/L+L-glutamine 500mg/+ sucrose 20g/L+ agar 6.5g/L+ active carbon 2g/L+ inositol 4g/L; PH=5.8;
Five, the growth of somatic embryo and plant regeneration
Callus with globular embryo is inoculated in medium and carries out body embryonic development, complete and turn embryo process, obtain cotyledonary embryos; Maturation induction is carried out to the cotyledonary embryos obtained simultaneously, obtains mature embryo, then carry out sprouting, plant regeneration and transplanting seedling.
Cones of Pinus koraiensis in present embodiment is collected in Wei He State Forestry Administration, P.R. China of Heilongjiang Province green hill tree seed orchard.
Present embodiment passes through the rational proportion of each component in medium and suitable condition of culture, make Korean pine when carrying out somatic embryo and occurring, frequency of embryonic callus induction and turn embryo rate and be obtained for raising, frequency of embryonic callus induction can reach 33.3%, present embodiment finds that inositol plays an important role to the induction of globular embryo and later stage body embryo maturation, by adding inositol, embryo callus subculture brownization degree is reduced, and more contribute to globular embryo and transform to mature embryo, turn embryo rate be do not add inositol 6-8 doubly.
Embodiment two: present embodiment and embodiment one unlike: the method being in the cones of Pinus koraiensis sterilizing of cleavage polyembryony phase in step one is: cleaned by cones of Pinus koraiensis, then to prune fruit scale, be the ethanol postincubation 3 ~ 5min of 70% again by mass concentration, then use aseptic water washing 2 ~ 4 times, then soak 20 ~ 30min with mass concentration 10% clorox.Other is identical with embodiment one.
Embodiment three: present embodiment and embodiment one or two unlike: the method for sterilizing again in step one is: be the hydrogen peroxide process 8min of 3% by mass concentration, aseptic water washing 2 ~ 4 times.Other is identical with embodiment one or two.
Embodiment four: one of present embodiment and embodiment one to three are unlike light culture 60d in step 2.Other is identical with one of embodiment one to three.
Embodiment five: one of present embodiment and embodiment one to four unlike: in step 3, the medium of squamous subculture is DCR medium+agar 6.5g/L+L-glutamine 500mg/L+ sucrose 30g/L+2,4-D0.5g/L+6-BA0.1g/L+ acid hydrolyzed casein 0.5g/L; PH=5.8.Other is identical with one of embodiment one to four.
Embodiment six: one of present embodiment and embodiment one to five unlike: in step 3, squamous subculture is under the condition of 23 ~ 25 DEG C, light culture 15d.Other is identical with one of embodiment one to five.
Embodiment seven: one of present embodiment and embodiment one to six are unlike light culture 12d in step 4.Other is identical with one of embodiment one to six.
Embodiment eight: one of present embodiment and embodiment one to seven are unlike light culture 70d in step 5.Other is identical with one of embodiment one to seven.
Embodiment nine: one of present embodiment and embodiment one to eight unlike: in step 5, body embryo Maturation induction medium is DCR medium+agar 7.1g/L+L-glutamine 500mg/L+ sucrose 60g/L+ abscisic acid 20mg/L+PEG400050g/L+IBA0.2mg/L+ active carbon 2g/L; PH=5.8.Other is identical with one of embodiment one to eight.
Embodiment ten: one of present embodiment and embodiment one to nine unlike: step five body constituents embryo Maturation induction is under the condition of 23 ~ 25 DEG C, light culture 60 ~ 80d, and every 3 ~ 4 weeks subcultures are once.Other is identical with one of embodiment one to nine.
The following examples will be further described the present invention, but not thereby limiting the invention.
Embodiment 1: the present embodiment improves Korean pine frequency of embryonic callus induction and turns the method for embryo rate, carries out according to the following steps:
One, the sterilizing of material
The cones of Pinus koraiensis being in the cleavage polyembryony phase is cleaned, then to prune fruit scale, be the ethanol postincubation 2min of 70% again by mass concentration, then aseptic water washing is used 3 times, soak 25min with mass concentration 10% clorox again, under superclean bench, peelling off seed coat afterwards, is then the hydrogen peroxide process 8min of 3% by mass concentration, aseptic water washing 3 times, obtains the zygotic embryo after sterilizing;
Two, embryo callus subculture induction
Zygotic embryo after sterilizing step one obtained is inoculated in medium, under the condition of 25 DEG C, light culture 60d, obtain embryo callus, wherein medium is DCR medium+agar 6.5g/L+L-glutamine 500mg/L+ sucrose 35g/L+NAA2mg/L+6-BA1.5mg/L+ acid hydrolyzed casein 0.8g/L; Medium pH=5.8; As shown in Figure 1, frequency of embryonic callus induction can reach 33.3% to the photo of the embryo callus of this step.
Three, the subculture of callus keeps and propagation
Embryo callus step 2 obtained carries out squamous subculture, under the condition of 25 DEG C, light culture 15d, obtain the callus after breeding, wherein the medium of squamous subculture is DCR medium+agar 6.5g/L+L-glutamine 500mg/L+ sucrose 30g/L+2,4-D0.5g/L+6-BA0.1g/L+ acid hydrolyzed casein 0.5g/L; PH=5.8;
Four, the induction of proembryo
Callus after propagation is inoculated into the induction carrying out proembryo in medium, and under the condition of 25 DEG C, light culture 12d, obtains the callus with globular embryo; Wherein medium is DCR medium+acid hydrolyzed casein 500mg/L+L-glutamine 500mg/+ sucrose 20g/L+ agar 6.5g/L+ active carbon 2g/L+ inositol 4g/L; PH=5.8; Generate the photo of early stage proembryos in this step process as shown in Figure 2, the photo of globular embryo as shown in Figure 3.
Five, the growth of somatic embryo
Callus with globular embryo is inoculated in medium and carries out body embryonic development, complete and turn embryo process; Maturation induction is carried out to the cotyledonary embryos obtained simultaneously, under the condition of 25 DEG C, light culture 60 ~ 80d, every 3 ~ 4 weeks subcultures once, obtain mature embryo, wherein medium is DCR medium+agar 7.1g/L+L-glutamine 500mg/L+ sucrose 60g/L+ abscisic acid 20mg/L+PEG400050g/L+IBA0.2mg/L+ active carbon 2g/L; PH=5.8; The photo of this step embryonic development as shown in Figure 4 and Figure 5.
Six, the sprouting of somatic embryo and plant regeneration
The mature somatic embryo level of induction is placed on germination medium DCR and sprouts, additional 20g/L sucrose, Glu 500mg/L, acid hydrolyzed casein 2g/L, first light culture 6d, then at illuminance 2000lx, 16/8h light/light culture 14d, carries out sprouting to mature somatic embryo and cultivates; Then adopt WPM minimal medium, additional 1.0mg/LIBA, 0.2mg/LNAA, 20g/L sucrose, 0.1g/L inositol and 6.5/L agar, illuminance 20001x, the light light culture of 16/8 hour 1 month, carries out somatic embryo plant regeneration, obtains regeneration plant;
Seven, the transplanting of regeneration plant
Transplant after more than plant root lengthen by 2 cm to be regenerated, before transplanting, sealed membrane is opened by 7d gradually, and make seedling adapt to extraneous environment, transplanting adopting volume ratio to be 1:3:1 perlite, Nutrition Soil and vermiculite is matrix, carries out transplanting and cultivates, namely complete.
Cones of Pinus koraiensis in the present embodiment is collected in Wei He State Forestry Administration, P.R. China of Heilongjiang Province green hill tree seed orchard.
The present embodiment makes embryo callus subculture brownization degree reduce by adding inositol, and more contributes to globular embryo and transform to mature embryo, conversion ratio be do not add inositol 6-8 doubly.
Known by above embodiment, the present invention passes through the rational proportion of each component in medium and suitable condition of culture, make Korean pine when carrying out somatic embryo and occurring, frequency of embryonic callus induction and turn embryo rate and be obtained for raising, frequency of embryonic callus induction can reach 33.3%, the present invention finds that inositol plays an important role to the induction of globular embryo and later stage body embryonic development and maturation, by adding inositol, embryo callus subculture brownization degree is reduced, and more contributing to the conversion of globular embryo, conversion ratio is 6-8 times that does not add inositol.
The above; be only the present invention's preferably embodiment, but protection scope of the present invention is not limited thereto, is anyly familiar with those skilled in the art in the technical scope that the present invention discloses; the change that can expect easily or replacement, all should be encompassed within protection scope of the present invention.
Claims (10)
1. improve Korean pine frequency of embryonic callus induction and the method turning embryo rate, it is characterized in that the method is carried out according to the following steps:
One, the sterilizing of material
By be in the cleavage polyembryony phase cones of Pinus koraiensis sterilizing after, peel off seed coat, again sterilizing, obtain the zygotic embryo after sterilizing;
Two, embryo callus subculture induction
Zygotic embryo after sterilizing step one obtained is inoculated in medium, under the condition of 23 ~ 25 DEG C, light culture 58 ~ 65d, obtain embryo callus, wherein medium is DCR medium+agar 6.5g/L+L-glutamine 500mg/L+ sucrose 35g/L+NAA2mg/L+6-BA1.5mg/L+ acid hydrolyzed casein 0.8g/L; Medium pH=5.8;
Three, the subculture of callus keeps and propagation
Embryo callus step 2 obtained carries out squamous subculture, obtains the callus after breeding;
Four, the induction of proembryo
Callus after propagation is inoculated into the induction carrying out proembryo in medium, and under the condition of 23 ~ 25 DEG C, light culture 7 ~ 15d, obtains the callus with globular embryo; Wherein medium is DCR medium+acid hydrolyzed casein 500mg/L+L-glutamine 500mg/+ sucrose 20g/L+ agar 6.5g/L+ active carbon 2g/L+ inositol 4g/L; PH=5.8;
Five, the growth of somatic embryo and plant regeneration
Callus with globular embryo is inoculated in medium and carries out body embryonic development, obtain cotyledonary embryos; Maturation induction is carried out to the cotyledonary embryos obtained simultaneously, obtains mature embryo, then carry out sprouting, plant regeneration and transplanting seedling.
2. a kind of method improving Korean pine frequency of embryonic callus induction and turn embryo rate according to claim 1, it is characterized in that the method for the cones of Pinus koraiensis sterilizing being in the cleavage polyembryony phase in step one is: cleaned by cones of Pinus koraiensis, then to prune fruit scale, be the ethanol postincubation 3 ~ 5min of 70% again by mass concentration, then use aseptic water washing 2 ~ 4 times, then soak 20 ~ 30min with mass concentration 10% clorox.
3. a kind of method improving Korean pine frequency of embryonic callus induction and turn embryo rate according to claim 1, is characterized in that the method for sterilizing again in step one is: be the hydrogen peroxide process 8min of 3% by mass concentration, aseptic water washing 2 ~ 4 times.
4. a kind of method improving Korean pine frequency of embryonic callus induction and turn embryo rate according to claim 1, is characterized in that light culture 60d in step 2.
5. a kind of method improving Korean pine frequency of embryonic callus induction and turn embryo rate according to claim 1, it is characterized in that the medium of squamous subculture in step 3 is DCR medium+agar 6.5g/L+L-glutamine 500mg/L+ sucrose 30g/L+2,4-D0.5g/L+6-BA0.1g/L+ acid hydrolyzed casein 0.5g/L; PH=5.8.
6. a kind of method improving Korean pine frequency of embryonic callus induction and turn embryo rate according to claim 1, is characterized in that in step 3, squamous subculture is under the condition of 23 ~ 25 DEG C, light culture 15d.
7. a kind of method improving Korean pine frequency of embryonic callus induction and turn embryo rate according to claim 1, is characterized in that light culture 12d in step 4.
8. a kind of method improving Korean pine frequency of embryonic callus induction and turn embryo rate according to claim 1, is characterized in that light culture 70d in step 5.
9. a kind of method improving Korean pine frequency of embryonic callus induction and turn embryo rate according to claim 1, is characterized in that in step 5, body embryo Maturation induction medium is DCR medium+agar 7.1g/L+L-glutamine 500mg/L+ sucrose 60g/L+ abscisic acid 20mg/L+PEG400050g/L+IBA0.2mg/L+ active carbon 2g/L; PH=5.8.
10. a kind of method improving Korean pine frequency of embryonic callus induction and turn embryo rate according to claim 1, it is characterized in that step five body constituents embryo Maturation induction is under the condition of 23 ~ 25 DEG C, light culture 60 ~ 80d, every 3 ~ 4 weeks subcultures once.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510771278.3A CN105432465B (en) | 2015-11-12 | 2015-11-12 | A kind of method for improving Korean pine frequency of embryonic callus induction and turning embryo rate |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510771278.3A CN105432465B (en) | 2015-11-12 | 2015-11-12 | A kind of method for improving Korean pine frequency of embryonic callus induction and turning embryo rate |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105432465A true CN105432465A (en) | 2016-03-30 |
| CN105432465B CN105432465B (en) | 2017-08-25 |
Family
ID=55543462
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510771278.3A Expired - Fee Related CN105432465B (en) | 2015-11-12 | 2015-11-12 | A kind of method for improving Korean pine frequency of embryonic callus induction and turning embryo rate |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105432465B (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106135005A (en) * | 2016-08-26 | 2016-11-23 | 李军 | A kind of little Semen Pini Tissue Culture Regeneration System construction method |
| CN108668899A (en) * | 2018-05-31 | 2018-10-19 | 广西壮族自治区林业科学研究院 | A method of improving masson pine body embryo inductivity |
| CN110004176A (en) * | 2019-04-12 | 2019-07-12 | 东北林业大学 | The construction method of hybrid larch genetic conversion system |
| CN110012835A (en) * | 2019-04-12 | 2019-07-16 | 江西省林业科学院 | A kind of method of wet-land pine tree embryonic callus induction and proliferation |
| CN110301353A (en) * | 2019-07-10 | 2019-10-08 | 广西壮族自治区林业科学研究院 | Method masson pine embryo callus subculture proliferation and maintain culture |
| CN112931215A (en) * | 2021-03-31 | 2021-06-11 | 东北林业大学 | Method for improving induction rate of embryonic callus of Korean pine |
| CN113068617A (en) * | 2021-05-14 | 2021-07-06 | 东北林业大学 | Method for improving germination rate of Korean pine somatic embryos through low-temperature treatment |
| CN113736821A (en) * | 2021-09-06 | 2021-12-03 | 东北林业大学 | Genetic transformation method for Korean pine embryonic callus |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4060933A (en) * | 1976-12-20 | 1977-12-06 | Gte Laboratories Incorporated | Tissue culture technique utilizing a specific light source |
| CN1476750A (en) * | 2002-04-09 | 2004-02-25 | 韦尔豪泽公司 | Method for producing zygotic appearance cotyledon type pine embryo in high yield by using culture medium containing diose and glucose |
| CN102577956A (en) * | 2012-02-21 | 2012-07-18 | 南京林业大学 | Pinus thunbergii cell embryogenesis and plant regeneration method |
-
2015
- 2015-11-12 CN CN201510771278.3A patent/CN105432465B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4060933A (en) * | 1976-12-20 | 1977-12-06 | Gte Laboratories Incorporated | Tissue culture technique utilizing a specific light source |
| CN1476750A (en) * | 2002-04-09 | 2004-02-25 | 韦尔豪泽公司 | Method for producing zygotic appearance cotyledon type pine embryo in high yield by using culture medium containing diose and glucose |
| CN102577956A (en) * | 2012-02-21 | 2012-07-18 | 南京林业大学 | Pinus thunbergii cell embryogenesis and plant regeneration method |
Non-Patent Citations (1)
| Title |
|---|
| 曹焱: ""红松合子胚愈伤组织诱导及不定芽发生的研究"", 《中国优秀硕士学位论文全文数据库农业科技辑》 * |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106135005A (en) * | 2016-08-26 | 2016-11-23 | 李军 | A kind of little Semen Pini Tissue Culture Regeneration System construction method |
| CN108668899A (en) * | 2018-05-31 | 2018-10-19 | 广西壮族自治区林业科学研究院 | A method of improving masson pine body embryo inductivity |
| CN108668899B (en) * | 2018-05-31 | 2021-04-20 | 广西壮族自治区林业科学研究院 | Method for improving induction rate of masson pine somatic embryos |
| CN110004176A (en) * | 2019-04-12 | 2019-07-12 | 东北林业大学 | The construction method of hybrid larch genetic conversion system |
| CN110012835A (en) * | 2019-04-12 | 2019-07-16 | 江西省林业科学院 | A kind of method of wet-land pine tree embryonic callus induction and proliferation |
| CN110004176B (en) * | 2019-04-12 | 2023-03-10 | 东北林业大学 | Construction method of hybrid larch genetic transformation system |
| CN110301353A (en) * | 2019-07-10 | 2019-10-08 | 广西壮族自治区林业科学研究院 | Method masson pine embryo callus subculture proliferation and maintain culture |
| CN112931215A (en) * | 2021-03-31 | 2021-06-11 | 东北林业大学 | Method for improving induction rate of embryonic callus of Korean pine |
| CN113068617A (en) * | 2021-05-14 | 2021-07-06 | 东北林业大学 | Method for improving germination rate of Korean pine somatic embryos through low-temperature treatment |
| CN113736821A (en) * | 2021-09-06 | 2021-12-03 | 东北林业大学 | Genetic transformation method for Korean pine embryonic callus |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105432465B (en) | 2017-08-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105432465A (en) | Method for improving embryonic callus induction rate and embryo transfer rate of pinus koraiensis | |
| Singh et al. | A simple regeneration protocol from stem explants of Jatropha curcas—a biodiesel plant | |
| CN104663450A (en) | Tissue culture and rapid propagation method for Acer rubrum 'Brandywine' | |
| CN102239805A (en) | Tissue-culture quick breeding method for iris tectorum | |
| CN102812905A (en) | Blueberry tender stem tissue culturing and rapid propagation process | |
| CN111616052A (en) | Rapid propagation and sugar-free rooting culture method and application of apple rootstock catalpa bungei | |
| CN112273231A (en) | Method for inducing proliferation of asparagus cochinchinensis cluster buds and plant regeneration | |
| CN108739370B (en) | Method for rapid propagation by utilizing mature lotus embryos | |
| CN105265316B (en) | A kind of allium plateau rapid propagation method | |
| CN104604677A (en) | Tissue culture propagation method for reducing tissue culture browning rate of traxacum koksaghyz | |
| Devi et al. | In vitro propagation of Arundinaria callosa Munro-an edible bamboo from nodal explants of mature plants | |
| US7932086B2 (en) | Commercially viable process for in vitro mass culture of Jatropha curcas | |
| CN105918128B (en) | A kind of American red-maple rapid propagation method | |
| Purohit et al. | Micropropagation of Achras sapota through enhanced axillary branching | |
| CN103299901A (en) | In-vitro rapid proliferation method of Masui dauphine fig | |
| CN112470931A (en) | Breeding method for axillary bud tissue culture of Jingning magnolia | |
| CN105613288A (en) | Construction method of rapid Euonymus japonicus L.f. aureo-marginatus Rehd propagation system | |
| CN112425506A (en) | Tissue culture rapid propagation method of artemisia anomala | |
| CN102630564B (en) | Tissue culture and rapid propagation method of salt-tolerant field mint | |
| EP1817956A2 (en) | Commercially viable process for in-vitro mass culture of jatropha curcas | |
| Deb et al. | Search of alternative substratum for agar in plant tissue culture | |
| CN103907535B (en) | Method for obtaining large number of bambusa glaucophylla regeneration plants through tissue culture | |
| North et al. | Effects of various media compositions on the'in vitro'germination and discoloration of immature embryos of bird of paradise ('Strelitzia reginae') | |
| CN104686336A (en) | Tissue culture rapid propagation method of ailanthus altissima | |
| CN112106664B (en) | Sterile germination and rapid propagation method for michelia spectabilis seeds |
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 |
Granted publication date: 20170825 Termination date: 20171112 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |