CN112753582B - Method for sterilizing and rapidly proliferating stem segments of aleurites montana - Google Patents
Method for sterilizing and rapidly proliferating stem segments of aleurites montana Download PDFInfo
- Publication number
- CN112753582B CN112753582B CN202110161400.0A CN202110161400A CN112753582B CN 112753582 B CN112753582 B CN 112753582B CN 202110161400 A CN202110161400 A CN 202110161400A CN 112753582 B CN112753582 B CN 112753582B
- Authority
- CN
- China
- Prior art keywords
- culture
- stem
- aleurites montana
- buds
- aleurites
- 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.)
- Active
Links
Images
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
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/40—Afforestation or reforestation
Abstract
The invention provides a method for disinfecting and quickly proliferating aleurites montana stem segments. Comprises the processes of disinfection of explants, axillary bud induction, rooting culture and the like. The invention overcomes the problems of high pollution rate and difficult culture of the primary culture of the stem section of the aleurites montana, has the advantages of low explant pollution rate, high propagation rate and short culture period, provides a way for rapidly propagating excellent aleurites montana aseptic seedlings, and provides possibility for industrial seedling culture of the aleurites montana and a later transgenic technology.
Description
Technical Field
The invention belongs to the field of aleurites montana tissue culture, and particularly relates to a method for disinfecting and rapidly proliferating aleurites montana stem sections.
Background
The aleurites montana (Vernicia montana) is a deciduous tree of Vernicia (Vernicia Lour.) of Euphorbiaceae (Euphorbiaceae), is a typical southern Asia tropical tree species, and has cultivation distribution in northern latitude 22 degrees 15 'to 34 degrees 30' and east longitude 99 degrees 40 'to 122 degrees 07', and main cultivation areas are Fujian, Guangdong, Guangxi, Jiangxi, Hunan and other provinces. The oil content of the dried seed kernels of the aleurites montana is 60-70%, one of the cultivation purposes is to extract oil from seeds and develop the oil into a biomass energy forest. The composite material has good fast-growing and cold-resistant quality, also has good development potential when being used as a tree species, and particularly has freezing resistance obviously superior to that of eucalyptus in an extremely low temperature hazard area. As the deciduous tree species are adopted, the mixed planting of forest farmers can be carried out during the scattered planting, the forest land can be fully utilized, and the main ecological planting is realized. In addition, because of easy planting, the ecological public welfare forest can be used as a good tree species (Zhongshou Wang 2011) for updating. The aleurites montana as a large arbor grows slowly in natural environment and is mainly planted by direct sowing, and the growth period is long. Tissue culture is an important way for quickly raising seedlings and shortening the growth cycle, and has wide commercial prospect. At present, few reports about tissue culture of aleurites montana are reported, the tissue culture of the aleurites montana is still in a starting stage, mainly because the problem that an explant is easy to pollute during primary culture exists in a stem section with buds of the aleurites montana in a tissue culture process, and a virus-free seedling is not cultured through the tissue culture until now.
At present, no tissue culture of aleurites montana appears, and only some patents on the tissue culture of the aleurites fordii are as follows:
201310271826.7 method for regenerating tung tree leaf
201410326807.4 method for inducing hypocotyl callus of tung tree and regenerating plant efficiently
201410327567.X method for directly regenerating plants efficiently from petioles of tung tree
201910934260.9 method for promoting regeneration of tung tree stem with bud by physical assistance
201610089435.7 method for directly inducing plant regeneration by using hypocotyl of Aleurites fordii as explant
201010252415.X agrobacterium tumefaciens-mediated gene transformation method of jatropha curcas
201710077542.2 method for rooting tissue culture seedling of tung tree in bottle
Compared with the patents, the invention has the advantages of simple and convenient operation, short culture period, simple formula of the culture medium and low required cost, and is particularly suitable for the rapid propagation of aleurites montana.
Disclosure of Invention
The invention aims to provide a method for disinfecting annual semi-lignified stem segments with buds of aleurites montana, aiming at the problems that the sterilization of the aleurites montana explant is difficult at present and the technology for obtaining a regenerated plant through tissue culture is insufficient. The research on the aspect of aleurites montana tissue culture is less, and the method is compared and analyzed with the tissue culture of the stem segment with the bud of the aleurites fordii of the same genus, has simple operation and good disinfection effect, omits the step of subculture proliferation, obviously shortens the growth and seedling period, has high axillary bud induction rate and robust root growth, and improves the possibility for the subsequent industrialized seedling and transgenic technology.
The primary object of the present invention is achieved as follows.
A method for disinfecting an explant of a stem segment with buds of aleurites montana comprises the following steps:
(1) obtaining an explant: planting Aleurites montana seedlings indoors or on the roof, and taking the current-year semi-lignified stem with buds as explant materials when the seedlings grow to 25-35 cm;
(2) treating explant, cutting annual semi-lignified stem segment with bud of Aleurites montana into 3-4cm long single-bud stem segment, washing with washing powder for 1-2 times, and washing with running water for at least 20 min;
(3) and (3) disinfection of explants: soaking the semi-lignified stem with buds washed by running water with 75% alcohol for 1min, replacing with new 75% alcohol for 1min, washing with sterile water for 3 times, sterilizing with 0.1% mercuric chloride for 45min, replacing mercuric chloride every 15min for 3 times, and finally washing with sterile water for 5-6 times.
According to the invention, a large number of experiments at the early stage show that the stem with buds of aleurites montana is very difficult to disinfect, various disinfection and sterilization modes are tried, the pollution rate is always high, an effective solution is finally obtained through multiple attempts, and the effect is very obvious after multiple experiments.
The second purpose of the invention is to provide a method for directly inducing the annual semi-lignified bud stem explant of aleurites montana to generate a complete plant, the method directly induces axillary buds by sterilizing the bud stem explant, and the method has high induction rate and good growth condition. The invention mainly explores in the aspects of axillary bud induction (primary culture), rooting culture and the like to form a set of tissue culture and rapid propagation technical system suitable for aleurites montana, and lays a foundation for industrial seedling culture and later transgenic technology of aleurites montana.
A method for inducing the annual semi-lignified budded stem of aleurites montana to rapidly proliferate, which comprises the following steps:
(1) axillary bud induction: inoculating sterilized stem segment of Aleurites montana with single bud to 1/2MS +0.1-2.0 mg/L6-BA +0.1mg/L IAA +0-0.2mg/L GA3The culture medium is subjected to induced culture, and the culture is carried out under the condition of illumination after dark culture;
(2) rooting culture: cutting off strong axillary buds obtained by stem segment differentiation, and inoculating the strong axillary buds into a culture medium of 1/2MS +0.5-2.0mg/L IBA +0-1.0mg/L NAA for rooting culture.
The medium used for induction culture in step (1) is preferably 1/2MS +2.0 mg/L6-BA +0.1mg/L IAA.
The culture medium used in the rooting culture in step (2) is preferably 1/2MS +2.0mg/L IBA.
Culturing in dark for 36h during axillary bud induction in the step (1), and culturing for 28-30 days under the illumination condition.
The culture temperature in the step (1) is 29 +/-1 ℃, the illumination culture intensity is 2100-.
According to the method for rapidly propagating aleurites montana, 30g/L of sucrose and 7g/L of agar are added into an induction culture medium, and the pH is adjusted to 5.5-5.8; adding sucrose 20g/L and agar 7g/L to the rooting culture medium, and adjusting pH to 5.5-5.8.
According to the method for rapidly propagating the annual semi-lignified stem segment with buds of the aleurites montana, the sterilized stem segment with single buds of the aleurites montana is sterilized by the method.
In addition, studies on tissue culture of Aleurites montana are rare. The differences and advantages of the studied and the present invention are summarized as follows:
1. the invention adopts the stem section with buds of the aleurites montana to successfully cultivate the tissue culture seedling of the aleurites montana, and has the characteristics of short growth period, good effect of inducing the axillary buds by primary culture and strong growth of the tissue culture seedling.
2. The invention utilizes a series of processes of disinfection treatment, primary culture, rooting culture, hardening-seedling transplantation and the like of the stem section with buds of the aleurites montana, aims to establish a high-efficiency and stable tissue culture and rapid propagation technical system of the aleurites montana and provides a technical basis for industrial seedling culture and transgenosis of the aleurites montana.
3. In the early stage experiment, the stem of aleurites montana is used for axillary bud induction and rooting culture, and although a plant with vigorous growth is obtained, the pollution rate and browning rate are high, and sterile tissue culture seedlings are difficult to obtain. Through a large number of comparative tests, the optimal disinfection method suitable for disinfecting the explant of the stem segment with the bud of the aleurites montana is finally obtained.
4. The method utilizes the strong axillary buds induced by the primary culture of the stem segments with the buds of the aleurites montana to directly induce rooting, has good rooting effect, vigorous seedling growth and greatly shortened culture period.
5. The invention can obtain regeneration plants through induction culture, but the axillary buds obtained through primary culture induction have the problem of difficult rooting. The invention obtains a formula suitable for direct rooting culture after the induction of the axillary buds of the aleurites montana through a large number of experiments, and compared with the tissue culture of the stem segments with buds of the aleurites fordii belonging to the same genus, the invention is simple and easy to implement and convenient to operate.
6. Compared with tissue culture of tung tree stem with buds belonging to the same genus, the sterilization of the explant is one of the key points of the invention, the sterilization effect of the aleurites montana explant material is obvious, the pollution rate is well controlled, and important guarantee is provided for subsequent culture.
7. The other key point of the invention is that axillary buds of the stem segment with buds of the aleurites montana root directly take roots, people like the beautiful flowers of Tangchun and the like use the stem segment with buds of the aleurites fordii to carry out tissue culture, although complete plants are obtained, the experimental process is quite complicated, a series of steps such as primary culture, enrichment culture, strong bud culture, rooting culture and the like are needed, and 80-95 days are needed from the primary culture to the rooting culture. Meanwhile, the culture medium and the formula of the invention are simple, the operation is convenient, and in contrast, the invention has the characteristics of short period, simple and convenient operation, cost reduction, obvious effect and the like.
Drawings
FIG. 1 is a photograph of an initial stage of inoculation of a stem segment of Aleurites montana of the present invention;
FIG. 2 is a photograph of an initial stage of inoculation of a stem segment of Aleurites montana of the present invention;
FIG. 3 is a photograph of a tung tree stem section of the present invention which has been disinfected for an extended period of time to brown but remains contaminated;
FIG. 4 is a photograph showing the growth of axillary buds of a stem segment of Aleurites montana of the present invention, which still have been contaminated;
FIG. 5 shows the stem segments of Aleurites montana added with GA3The photograph of (a);
FIG. 6 shows the stem segments of Aleurites montana added with GA3Photograph of medium culture;
FIG. 7 is a photograph of the induction of axillary buds of Aleurites montana of the present invention for 25 days;
FIG. 8 is a photograph of axillary bud induction of Aleurites montana of the present invention for 30 days;
FIG. 9 is a photograph of an early stage of rooting culture of a stem segment of Aleurites montana according to the present invention;
FIG. 10 is a photograph of rooting a stem segment of Aleurites montana of the present invention;
FIG. 11 is a photograph of a rooting culture of a stem segment of Aleurites montana of the present invention for 24 days;
FIG. 12 is a photograph of a rooting culture of a stem segment of Aleurites montana of the present invention for 24 days;
FIG. 13 is a photograph of a sterile seedling of Aleurites montana cultured during the course of this experiment.
Detailed Description
The following examples are intended to further illustrate the invention without limiting it.
Example 1
The following operations are carried out in sequence:
1. and (3) explant disinfection treatment: in the morning of 4-6 months in 2019, selecting young and tender Aleurites montana stems with moderate thickness in the same year on the roof, spraying water, taking the stems back to a laboratory to remove leaves, washing the stems for 1-2 times by using a proper amount of washing powder, washing the stems for at least about 20min by using running water, washing the stems clean by using sterile water, placing the stems in a superclean bench, disinfecting the stems by using 75% of alcohol and 0.1% of mercury bichloride, and setting different time gradients as shown in Table 1. Continuously stirring with tweezers during the disinfection process, washing with sterile water for 5-6 times, sucking water on the surface of the stem segment with sterile filter paper, cutting off browned parts at two ends with a sterile blade, inoculating the single-bud stem segment into the culture medium, and counting the pollution rate and the browning rate;
TABLE 1 Disinfection Effect of different Disinfection treatments on Aleurites montana Stem segments
Finally, it was confirmed that the preferred sterilization method of the present invention is as follows:
(1) and (3) explant treatment, namely cutting the annual semi-lignified stem section with buds of aleurites montana into 3-4cm stem sections with single buds, washing for 1-2 times by using washing powder, and washing for more than 20min by using running water.
(2) Explant disinfection: soaking the treated semi-lignified stem with buds in 75% alcohol for 1min, soaking in 75% alcohol for 1min, washing with sterile water for 3 times, sterilizing with 0.1% mercuric chloride for 45min, changing mercuric chloride every 15min for 3 times, and washing with sterile water for 5-6 times.
2. Inoculating the sterilized explant into 1/2MS culture medium according to the hormone formula in Table 2 for axillary bud induction, culturing in dark for 36h, and culturing under illumination for 28-30d, wherein the highest induction rate can reach 84.32%. Adding 30g/L of sucrose, 7g/L of agar and 5.5-5.8 of pH; the culture temperature is 29 ℃, the illumination intensity is 2100-.
TABLE 2 Effect of different plant growth regulators on Primary culture of Aleurites montana Stem segments
Finally, the preferred axillary bud induction medium of the present invention was confirmed to be: 1/2MS +2.0 mg/L6-BA +0.1mg/L IAA.
3. The axillary buds obtained by primary culture are inoculated into 1/2MS culture medium according to the hormone formula in the table 3 for rooting induction, 20g/L of sucrose, 7g/L of agar and 5.5-5.8 of pH are added. The culture temperature is 29 ℃, the illumination intensity is 2100-.
TABLE 3 Effect of different hormone combinations on rooting of aleurites montana test-tube plantlets
Finally, the preferable rooting culture medium of the invention is confirmed to be: 1/2MS +2.0mg/L IBA.
The tung oil tree patents mentioned in the background art are numbered as D1, D2 and … … D7 from top to bottom, the invention is D8, and the time length from primary culture to complete plant formation is analyzed:
d1:201310271826.7 method for regenerating plants by tung tree leaves
D2:201410326807.4 method for inducing hypocotyl callus of tung tree and efficiently regenerating plant
D3:201410327567.X method for directly regenerating plants efficiently from tung tree petioles
D4:201910934260.9 method for physically assisting tung tree bud-carrying stem segment regeneration plant
D5:201610089435.7 method for directly inducing plant regeneration by using tung tree hypocotyl as explant
D6:201010252415.X Agrobacterium mediated Jatropha curcas gene transformation method
Method for rooting in D7:201710077542.2 tissue culture seedling bottle of tung tree
TABLE 4 Effect of different cultivation methods on the duration of cultivation
Because of the difference of varieties of the aleurites fordii and the aleurites montana, the aleurites montana explant adopts the primary generation or proliferation culture medium of the aleurites fordii and the rooting culture medium to replace the primary generation induction culture medium and the rooting culture medium to culture the stem section with the buds of the aleurites montana in the invention, which are not suitable, and the induction and rooting effects are extremely poor and far inferior to the culture medium in the invention.
In the above table, D4 is a patent before the subject group of the applicant, so that the technical scheme is completely understood, the scheme utilizes physical induction to promote the regeneration of the tung tree plants through more cultivation processes, and also needs the assistance of physical means, the cultivation time is far longer than that of the present invention, and the key is that the stem induction medium with buds, the rooting medium and the cultivation mode in the scheme are not suitable for the cultivation of the aleurites montana of the present invention, so that the present invention re-explores the cultivation scheme for the aleurites montana; d6 is a gene transformation method of tung tree mediated by agrobacterium, which has a big difference with the main direction of the invention and needs more culture process, so the culture time is longer. The data in the table show that the method has great advantages in culture time, the time required by the method for culturing the stem with the buds of the aleurites montana from the initial generation to the root growth is almost shortened by more than two times compared with other patents, the biggest characteristic of the method is fully shown, namely, the breeding period is shortened, the operation is simple and easy, the culture cost is reduced, and a foundation is provided for the development of industrial seedling culture and transgenic technology.
Claims (4)
1. A method for inducing the rapid proliferation of the annual semi-lignified stem with buds of aleurites montana is characterized by comprising the following steps:
(1) axillary bud induction: inoculating the disinfected aleurites montana stem with a single bud into a culture medium of 1/2MS +2.0 mg/L6-BA +0.1mg/L IAA for induction culture, and culturing under the condition of illumination after dark culture;
(2) rooting culture: cutting off strong axillary buds obtained by stem segment differentiation, and inoculating the strong axillary buds into a culture medium of 1/2MS +2.0mg/L IBA for rooting culture;
the method for disinfecting the explant of the stem with buds of aleurites montana comprises the following steps:
1) obtaining an explant: planting Aleurites montana seedlings indoors or on the roof, and taking the current-year semi-lignified stem with buds as explant materials when the seedlings grow to 25-35 cm;
2) processing explant, cutting the annual semi-lignified stem segment with bud of Aleurites montana into 3-4cm long single-bud stem segment, washing with washing powder for 1-2 times, and washing with running water for at least 20 min;
3) and (3) disinfection of explants: soaking the semi-lignified stem with buds washed by running water with 75% alcohol for 1min, replacing with new 75% alcohol for 1min, washing with sterile water for 3 times, sterilizing with 0.1% mercuric chloride for 45min, replacing mercuric chloride every 15min for 3 times, and finally washing with sterile water for 5-6 times.
2. The method according to claim 1, wherein the axillary bud is cultured under light for 28 to 30 days after culturing for 36 hours in the dark at the time of axillary bud induction in step (1).
3. The method as claimed in claim 1, wherein the culturing temperature in step (1) is 29. + -. 1 ℃, the illumination culturing intensity is 2100-.
4. The method of claim 1, wherein the induction medium is supplemented with sucrose at 30g/L, agar at 7g/L, and pH adjusted to 5.5-5.8; adding sucrose 20g/L and agar 7g/L to the rooting culture medium, and adjusting pH to 5.5-5.8.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110161400.0A CN112753582B (en) | 2021-02-05 | 2021-02-05 | Method for sterilizing and rapidly proliferating stem segments of aleurites montana |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110161400.0A CN112753582B (en) | 2021-02-05 | 2021-02-05 | Method for sterilizing and rapidly proliferating stem segments of aleurites montana |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112753582A CN112753582A (en) | 2021-05-07 |
| CN112753582B true CN112753582B (en) | 2022-06-03 |
Family
ID=75705107
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110161400.0A Active CN112753582B (en) | 2021-02-05 | 2021-02-05 | Method for sterilizing and rapidly proliferating stem segments of aleurites montana |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112753582B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110476816B (en) * | 2019-09-29 | 2022-10-14 | 西南大学 | Method for quickly obtaining grapevine seedlings |
| CN114680047B (en) * | 2022-05-31 | 2022-08-12 | 华南农业大学 | Tissue culture rapid propagation method taking field tree spinach stem as explant |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP5710155B2 (en) * | 2010-05-31 | 2015-04-30 | 一般財団法人電力中央研究所 | Plant cell culture medium and plant cell culture method using the same |
| CN106718940A (en) * | 2017-02-15 | 2017-05-31 | 唐春艳 | A kind of tissue culture and rapid propagation method suitable for tung oil tree |
| CN110476818B (en) * | 2019-09-29 | 2020-08-14 | 中南林业科技大学 | Method for physically assisting regeneration of plant from tung tree stem with bud |
| CN110476816B (en) * | 2019-09-29 | 2022-10-14 | 西南大学 | Method for quickly obtaining grapevine seedlings |
-
2021
- 2021-02-05 CN CN202110161400.0A patent/CN112753582B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN112753582A (en) | 2021-05-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109479715B (en) | Method for rapidly breeding hydrangea macrophylla endless summer by using tissue culture seedling leaves | |
| CN112753582B (en) | Method for sterilizing and rapidly proliferating stem segments of aleurites montana | |
| CN112273231A (en) | Method for inducing proliferation of asparagus cochinchinensis cluster buds and plant regeneration | |
| CN114051932A (en) | Method for establishing efficient rapid propagation system by taking stem segments with axillary buds of tea trees as explants | |
| CN110558233B (en) | Method for disinfecting explant of stem segment with buds of black tiger and method for directly inducing aseptic buds to rapidly proliferate | |
| CN106538382B (en) | Method for establishing efficient eremochloa ophiuroides regeneration system by taking young ears as explants | |
| CN110583488A (en) | Method for establishing tissue culture rapid propagation technical system of new lycoris variety' pink | |
| CN102524065B (en) | High frequency regeneration method of Jatropha curcas | |
| CN112425506A (en) | Tissue culture rapid propagation method of artemisia anomala | |
| CN109601388B (en) | Tissue culture rapid propagation method of hybrid clematis | |
| CN110771505A (en) | Culture method of citral tonglu stem tissue | |
| CN112616675B (en) | Tissue culture and rapid propagation method for Zingiber dance | |
| CN112400696B (en) | Tissue culture method of evergreen common selfheal fruit-spike bamboo | |
| CN110771512B (en) | Efficient induction method of rabdosia lophanthide callus | |
| CN109526748B (en) | Tissue culture method for anthurium andraeanum inflorescence | |
| CN109906939B (en) | Pepper in-vitro regeneration method and culture medium used by same | |
| CN110583481A (en) | Method for inducing somatic embryogenesis and plant regeneration of Aralia elata | |
| CN112514796B (en) | Tissue culture rapid propagation method of new variety 'Xueli' of vertical branch type chionanthus retusus | |
| CN111264393B (en) | Method for rapidly breeding epimedium test-tube plantlets | |
| CN112400692B (en) | Tissue culture and rapid propagation method of Chinese flowering crabapple | |
| CN111887152B (en) | Method for establishing plant regeneration system of swallow flower bud growing way | |
| CN115644056B (en) | Industrial production method for nepenthes tissue culture | |
| CN116369196B (en) | Method for inducing abnormal urine and thinning callus and regenerating plant | |
| CN117084170B (en) | Tissue culture rapid propagation method for leaves of Bumarc | |
| CN110432151B (en) | Tissue culture and rapid propagation method of Chongyang wood |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |