CN110089432B - Sugar-free tissue culture seedling method for paper mulberry based on environmental factor regulation - Google Patents
Sugar-free tissue culture seedling method for paper mulberry based on environmental factor regulation Download PDFInfo
- Publication number
- CN110089432B CN110089432B CN201910400470.XA CN201910400470A CN110089432B CN 110089432 B CN110089432 B CN 110089432B CN 201910400470 A CN201910400470 A CN 201910400470A CN 110089432 B CN110089432 B CN 110089432B
- Authority
- CN
- China
- Prior art keywords
- sugar
- culture
- free
- seedlings
- tissue culture
- 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
- A01H4/001—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
-
- 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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/25—Greenhouse technology, e.g. cooling systems therefor
Abstract
A sugar-free paper mulberry tissue culture seedling method based on environmental factor regulation and control comprises the steps of inoculating paper mulberry explants into a sugar-free culture container, regulating and controlling environmental factors, carrying out proliferation and propagation and synchronous rooting culture to obtain sugar-free tissue culture seedlings, repeatedly proliferating the sugar-free tissue culture seedlings for multiple times under the same culture condition, and directly transferring the sugar-free tissue culture seedlings into a seedling culture medium to delay seedlings after production demand is met, so that the whole seedling culture process is completed. The method can effectively improve the plant growth vigor and the rooting quality of the paper mulberry tissue culture seedlings under the production environment of non-strict disinfection and sterilization, reduce the pollution death rate, reduce the transplanting and seedling hardening procedures, shorten the seedling culture period, improve the labor productivity and reduce the production cost.
Description
Technical Field
The invention relates to a technology in the field of plant planting, in particular to a sugar-free tissue culture seedling method for broussonetia papyrifera based on environmental factor regulation.
Background
At present, paper mulberry (Broussonetia papyrifera) is mainly propagated by means of seeds, cuttings and plant tissue culture. The progeny of the seeding propagation can generate character separation, the good characteristics of the strain are not easy to maintain, the cutting propagation period is long, and the survival rate is low, so the high-quality paper mulberry seedling is basically produced by adopting a plant tissue culture method. However, in the tissue culture production, except for the inherent easy pollution, complex operation, poor ventilation and many abnormalities in plant morphology and physiological functions of the traditional tissue culture, the endophytes of the paper mulberry are difficult to completely remove, and the problems of high tissue culture pollution rate and low seedling rate are aggravated. The traditional paper mulberry tissue culture process has long growth period, weak growth vigor, difficult rooting, low survival rate, poor root development quality and long hardening-seedling period, and seriously restricts the industrial development of excellent paper mulberry varieties.
The search of the prior art shows that Chinese patent No. CN109122322A, published (published) Japanese 2019.01.04, discloses a method and a device for tissue culture and rapid propagation of catalpa bungei, wherein a sugar-free culture medium is adopted to culture catalpa bungei tissue culture seedlings, carbon dioxide is used as a carbon source instead of sugar, and the catalpa bungei tissue culture seedlings grow by autotrophy through photosynthesis. However, the organic matter component is used in the technology, so that the effective control of sterility or mixed bacteria can be ensured only by achieving extremely high cleanliness in the culture process, and the normal growth of plants is not influenced. But there is no corresponding filtration, bacteriostasis or sterilization measure in the system. So that the practical production is difficult to apply.
Chinese patent publication No. CN102668990A discloses (announced) japanese 2012.09.19, which discloses a plant tissue culture method and apparatus, under relatively aseptic environment, 3.4-5.5g of agar powder is weighed to prepare 1L of culture medium, agar is heated to be fully dissolved, then various elements except sucrose are added according to the culture stage, the pH value of the culture medium is adjusted to 5.6-5.8, and the sugar-free culture medium is obtained by constant volume and split charging; cleaning an inoculation table top and hands by using alcohol with the mass percentage concentration of 75% in an inoculation room, putting a sugar-free culture medium into a carbon dioxide gas fertilizer culture bottle, inoculating a plant explant onto the sugar-free culture medium, putting the carbon dioxide gas fertilizer into the carbon dioxide gas fertilizer culture bottle, and sealing the carbon dioxide gas fertilizer culture bottle; the dosage of the carbon dioxide gas fertilizer is 0.5-1.0g/250 ml. Carbon dioxide gas is used for replacing sucrose in the culture medium, so that a carbon source required by photosynthesis is provided for plants, and meanwhile, the bacteriostatic agent is added into the culture medium to inhibit the pollution of microorganisms, so that multiple effects of bacteriostasis, sterilization, simplification of operation and improvement of tissue culture seedling quality are achieved. However, the invention needs a specially designed carbon dioxide gas fertilizer culture bottle, and the carbon dioxide gas fertilizer must be synchronously put in each bottle during inoculation, so the operation is complicated, and the workload is extremely high; ② adding bacteriostatic agent into the culture medium.
Disclosure of Invention
Aiming at the defects of complex carbon dioxide treatment process and overhigh requirement on cultivation environment in the conventional sugar-free paper mulberry tissue culture, the invention provides a sugar-free paper mulberry tissue culture seedling method based on environmental factor regulation, which can effectively improve the plant growth vigor and the rooting quality of paper mulberry tissue culture seedlings in a common environment, reduce the pollution death rate, reduce the transplanting and seedling hardening procedures, shorten the seedling culture period, improve the labor productivity and reduce the production cost.
The invention is realized by the following technical scheme:
according to the invention, the paper mulberry explant is inoculated into a sugar-free culture container, environmental factors are regulated and controlled, proliferation and propagation and synchronous rooting culture are carried out, so as to obtain the sugar-free tissue culture seedling, the sugar-free tissue culture seedling is repeatedly proliferated for many times under the same culture condition, and after the production demand is reached, the sugar-free tissue culture seedling is directly transferred into a seedling culture substrate for seedling rejuvenation, so that the whole seedling culture process is completed.
The explant comprises plant parts obtained from nature, cultured with sugar or cultured without sugar, and ensures that each unit of explant has at least one leaf and one bud.
The plant part includes but is not limited to apical bud, stem segment or basal multiple bud.
The explant obtained from the nature is preferably subjected to disinfection treatment, and is subjected to surface disinfection for 5-15min by adopting but not limited to 0.01-1% potassium permanganate. The plant parts obtained by sugar culture or sugar-free culture do not need disinfection treatment.
The environmental regulation and control factors are as follows: setting illumination intensity at 0-5d sugar-free early stage of culture to 50-100 μmol. m -2 ·s -1 At the temperature of 18-25 ℃; setting the illumination intensity of 100-200 mu mol.m at the later period of 5-30d sugar-free culture -2 ·s -1 At the temperature of 18-35 ℃; the carbon dioxide concentration in the whole sugar-free culture process is as follows: the illumination period is 500-5000ppm, and the dark period is 380-1200 ppm.
The proliferation propagation and synchronous rooting culture means that explants are inoculated into a culture container of a sugar-free culture medium to be cultured for 15-30 days in a sugar-free way, and the sugar-free tissue culture seedlings of paper mulberry with complete roots, stems and leaves are obtained.
The sugar-free culture medium consists of inorganic salt, hormone, water and a substrate, and the components and the concentration in each liter are as follows: potassium nitrate 400-2800mg/L, ammonium nitrate 300-2400mg/L, magnesium sulfate heptahydrate 70-740mg/L, anhydrous potassium dihydrogen phosphate 30-340mg/L, calcium chloride dihydrate 80-660mg/L, disodium ethylene diamine tetraacetate 7.5-70mg/L, ferrous sulfate heptahydrate 5.5-93mg/L, manganese sulfate tetrahydrate 4.5-33mg/L, zinc sulfate 1.5-13mg/L, boric acid 1.5-10mg/L, potassium iodide 0.15-1.5mg/L, sodium molybdate 0.05-0.5mg/L, copper sulfate 0.005-0.04mg/L, cobalt chloride 0.005-0.04mg/L and acetyl Indole Butyric Acid (IBA)0.001-10 mg/L.
The matrix is selected from vermiculite, perlite, flower mud, sponge, sand, gravel and the like.
The pH value of the sugar-free culture medium is 5.8-6.2.
The culture container is made of transparent materials with holes.
The sugar-free culture environment factor setting comprises: the temperature is 18-35 deg.C, and the illumination intensity is 50-200 μmol · m -2 ·s -1 The number of illumination hours is 12-18 h/day, CO 2 The concentration is 380 mu mol & mol -1 -5000μmol·mol -1 。
The seedling revival means: transplanting the sugar-free paper mulberry tissue culture seedlings into a seedling raising shed with the temperature of 15-30 ℃, directly transplanting the paper mulberry tissue culture seedlings into a seedling raising matrix, shading and rejuvenating the seedlings for 2-5 days, and completing the whole seedling raising process after the plants recover to grow.
Technical effects
Compared with the prior art, the invention has the technical effects that:
1) the culture period is short. With CO 2 Sugar is replaced as the carbon source of the paper mulberry test-tube plantlet, the plant growth environment is dynamically adjusted and optimized, and the optimal CO is provided for the propagation and growth of the plantlet 2 The photosynthetic rate and growth of the plantlets are effectively promoted under the environmental conditions of concentration, illumination, temperature and the like; the loose porous air-permeable inorganic material is used as a rooting substrate, the rooting environment is optimized, the root system is early and fast grown, and the seedling strengthening and rooting culture stages are combined into a whole, so that the culture period of the paper mulberry is shortened.
2) The pollution rate is greatly reduced. In the traditional paper mulberry tissue culture, because endophytes are difficult to completely remove, sucrose in a culture medium is easy to cause large-area microbial pollution, and the batch death of seedlings is caused. Sugar-free tissue culture and utilization of CO 2 After sugar in the substitute culture medium is used as a carbon source, microorganisms lose the carbon source and cannot breed, and the pollution probability and the seedling death rate are greatly reduced.
3) The operation is simple. Because the pollution rate is greatly reduced, the operation on an ultra-clean workbench with 100-grade cleanliness in the Standard of drug production quality management Standard is not required in actual production, the relatively clean space can be used for the sugar-free culture production of the paper mulberry, and the production efficiency of workers is greatly improved after the complicated aseptic operation is omitted. Meanwhile, the large culture container is more convenient for production and operation, and can improve the utilization rate of the culture space.
4) Seedling hardening is not needed, and the transplanting survival rate is high. When the sugar tissue culture of the broussonetia papyrifera is carried out, the sugar is a nutrient source of the plants, the plants mainly obtain required nutrients in a heterotrophic mode, when the outdoor transplantation is carried out, the seedlings need to be subjected to self-nutrition mode conversion, and meanwhile, when the sugar tissue culture is carried out, the plants are in a small container closed culture environment, so that the organ functions of each part of the plants are different from those of outdoor photoautotrophic plants, and therefore, the sugar seedlings of the broussonetia papyrifera adapt to the external environment by a complicated seedling hardening-off procedure, and the transplanting survival rate is difficult to ensure; when sugar-free tissue culture of paper mulberry is carried out, the nutrition mode of the plant is autotrophy, each organ of the plantlet keeps normal tissue structure and air pore function by reasonable environment regulation in the culture container, and the tissue has higher light and light capacity and better external environment adaptation capacity during transplantation, thereby reducing the transplantation seedling hardening procedure and being very beneficial to improving the survival rate of transition seedlings.
Drawings
FIG. 1 is a photograph of sugar-free tissue culture seedlings in the examples;
FIG. 2 is a photograph of individual sugarless tissue culture seedlings of the examples.
Detailed Description
The embodiment relates to a sugar-free tissue culture and rapid propagation method of paper mulberry, which comprises the following steps:
step 1), selecting broussonetia papyrifera tissue culture seedlings with the plant height of 4-6cm, and inoculating the broussonetia papyrifera tissue culture seedlings into a sugar-free culture container. Vermiculite with specification of 2-5mm is used as culture medium, the culture medium adopts large amount, trace amount and ferric salt of MS basic culture medium, IBA 1.0mg/L is added, sugar and organic matter are not added, temperature is controlled at 25 + -2 deg.C, and illumination intensity is 60 + -10 μmol · m for five days before culture -2 ·s -1 Without supplying CO 2 (ii) a Then the illumination intensity is increased to 110 +/-10 mu mol.m -2 ·s -1 The number of illumination hours is 14 h/day, and the container is supplemented with CO every half hour during the illumination period 2 The concentration is 800 +/-50 mu mol & mol -1 The mixed air of (2) is used for 15min, and the sugar-free culture is carried out for 20 days under the environmental conditionsObtaining the sugar-free tissue culture seedling of paper mulberry with the plant height of 6-8 cm.
Step 2) the sugar-free tissue culture seedling of paper mulberry obtained in step 1) can be prolonged for culture for 30 days to obtain a rooting seedling with a height of 10-12cm, the plant is divided into stem segments with leaves of 3-4cm, the stem segments are inoculated into a sugar-free culture container in a clean room, vermiculite with a specification of 2-5mm is used as a culture medium, a large amount of trace iron salt and a large amount of trace iron salt of an MS basic culture medium are adopted, sugar and organic matter are not added, IBA is added in an amount of 0.5mg/L, the temperature is controlled to be 25 +/-2 ℃, and the illumination intensity is 60 +/-10 mu mol.m in five days before culture -2 ·s -1 Without supplying CO 2 (ii) a Then the illumination intensity is increased to 110 +/-10 mu mol.m -2 ·s -1， The number of illumination hours is 14 h/day, and the container is supplemented with CO every half hour during the illumination period 2 The concentration is 800 +/-50 mu mol & mol -1 The mixed air is mixed for 15min, and sugar-free culture is carried out for 20 days under the environmental conditions to obtain the paper mulberry sugar-free tissue culture seedling with the plant height of 6-8 cm.
And 3) transplanting the sugar-free tissue culture seedlings of the paper mulberry obtained in the step 1) and the step 2) into a seedling transplanting shed at the temperature of 25 +/-2 ℃, directly transplanting the seedlings into a seedling culture medium mixed by grass carbon and perlite (volume ratio is 3:1), shading and recovering seedlings for 3 days, and finishing the whole seedling culture process after the plants recover to grow.
Results and data: the conventional sugar-containing tissue culture is taken as a comparison example, and the differences of the planting quantity, the pollution rate, the survival rate, the culture period, the seedling hardening period and the like under the same culture area are obvious.
| Index \ culture type | Sugar-free culture | Comparative example |
| Number of single layer culture rack | 640a | 400b |
| Contamination ratio (%) | 0.13b | 36.7a |
| Survival rate (%) | 98.8a | 75.1b |
| Proliferative cycle (day) | 20b | 30a |
| Rooting cycle (sky) | 0b | 25a |
| Rooting percentage (%) | 99.2a | 83.5b |
| Seedling period (Tian) | 3b | 45a |
| Plant height (cm) | 7.8a | 3.5b |
| Number of blades (sheet) | 4.3a | 3.2b |
| Daily inoculum (strain/person, day) | 3260a | 2153b |
The foregoing embodiments may be modified in many different ways by one skilled in the art without departing from the spirit and scope of the invention, which is defined by the appended claims and not by the preceding embodiments, and all embodiments within their scope are intended to be limited by the scope of the invention.
Claims (1)
1. A sugar-free tissue culture seedling method of paper mulberry based on environmental factor regulation is characterized by comprising the following steps:
step 1), selecting broussonetia papyrifera tissue culture seedlings with the plant height of 4-6cm, and inoculating the broussonetia papyrifera tissue culture seedlings into a sugar-free culture container; vermiculite with specification of 2-5mm is used as culture medium, the culture medium adopts large amount, trace amount and ferric salt of MS basic culture medium, indoleacetic acid is added at 1.0mg/L, no sugar or organic matter is added, temperature is controlled at 25 + -2 deg.C, and illumination intensity is 60 + -10 μmol · m for five days before culture -2 ·s -1 Without supplying CO 2 (ii) a Then the illumination intensity is increased to 110 +/-10 mu mol.m -2 ·s -1 The illumination time is 14 h/day, and CO is supplemented into the container every half hour 2 The concentration is 800 +/-50 mu mol & mol -1 The mixed air is used for 15min, and sugar-free culture is carried out for 20 days under the environmental condition to obtain the paper mulberry sugar-free tissue culture seedlings with the plant height of 6-8 cm;
step 2) prolonging the culture time of the sugar-free tissue culture seedlings of paper mulberry obtained in the step 1) to 30 days to obtain rooting seedlings with the height of 10-12cm, dividing the plants into stem segments with leaves of 3-4cm, inoculating the stem segments into a sugar-free culture container in a clean room, taking vermiculite with the specification of 2-5mm as a culture medium, adopting a large amount of trace and ferric salts of an MS basic culture medium, adding no sugar and organic matters, adding 0.5mg/L of indoleacetic acid, controlling the temperature to be 25 +/-2 ℃, and controlling the illumination intensity to be 60 +/-10 mu mol.m for five days before culture -2 ·s -1 Without supplying CO 2 (ii) a Then the illumination intensity is increased to 110 +/-10 mu mol.m -2 ·s -1 14h light hoursSupplementing CO into the container every half an hour in day and illumination period 2 The concentration is 800 +/-50 mu mol & mol -1 The mixed air is used for 15min, and sugar-free culture is carried out for 20 days under the environmental condition to obtain the paper mulberry sugar-free tissue culture seedlings with the plant height of 6-8 cm;
and 3) transplanting the sugar-free tissue culture seedlings of the paper mulberry obtained in the step 1) and the step 2) into a seedling transplanting shed with the temperature of 25 +/-2 ℃, directly transplanting the seedlings into a seedling culture medium mixed with turf and perlite according to the volume ratio of 3:1, shading and reviving the seedlings for 3 days, and completing the whole seedling culture process after the plants recover to grow.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910400470.XA CN110089432B (en) | 2019-05-15 | 2019-05-15 | Sugar-free tissue culture seedling method for paper mulberry based on environmental factor regulation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910400470.XA CN110089432B (en) | 2019-05-15 | 2019-05-15 | Sugar-free tissue culture seedling method for paper mulberry based on environmental factor regulation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110089432A CN110089432A (en) | 2019-08-06 |
| CN110089432B true CN110089432B (en) | 2022-09-09 |
Family
ID=67448118
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910400470.XA Active CN110089432B (en) | 2019-05-15 | 2019-05-15 | Sugar-free tissue culture seedling method for paper mulberry based on environmental factor regulation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110089432B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110667908B (en) * | 2019-10-21 | 2022-08-02 | 北部湾大学 | Tissue culture seedling packaging method suitable for long-distance transportation |
| CN112772415B (en) * | 2021-01-27 | 2023-04-25 | 广州市农业科学研究院 | Method for rapid propagation of banana seedlings |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105532469A (en) * | 2016-01-12 | 2016-05-04 | 靳杏子 | Method for hybrid broussonetia papyrifera tissue culture |
| CN209845910U (en) * | 2019-04-03 | 2019-12-27 | 湖北省农业科学院果树茶叶研究所 | Three-dimensional rooting and seedling strengthening device for tissue culture seedlings of hybrid paper mulberry |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2001204283A (en) * | 2000-01-21 | 2001-07-31 | Nisshinbo Ind Inc | Apparatus for culturing plant body and method of culturing plant body using the same |
| CN106954551A (en) * | 2017-04-28 | 2017-07-18 | 上海离草科技有限公司 | Sugar Free Plant Tissue Culture special culture media |
| CN108633738A (en) * | 2018-05-15 | 2018-10-12 | 贵州务川科华生物科技有限公司 | A method of producing paper mulberry tissue-cultured seedling |
-
2019
- 2019-05-15 CN CN201910400470.XA patent/CN110089432B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105532469A (en) * | 2016-01-12 | 2016-05-04 | 靳杏子 | Method for hybrid broussonetia papyrifera tissue culture |
| CN209845910U (en) * | 2019-04-03 | 2019-12-27 | 湖北省农业科学院果树茶叶研究所 | Three-dimensional rooting and seedling strengthening device for tissue culture seedlings of hybrid paper mulberry |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110089432A (en) | 2019-08-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105075863B (en) | A kind of Paeonia papaveracea rapid propagation method | |
| CN101926259A (en) | Orchid germchit propagating method | |
| CN111616052A (en) | Rapid propagation and sugar-free rooting culture method and application of apple rootstock catalpa bungei | |
| CN110089432B (en) | Sugar-free tissue culture seedling method for paper mulberry based on environmental factor regulation | |
| CN111280056A (en) | Subculture breeding method of stingless pepper tissue culture seedlings | |
| CN111149704A (en) | Proliferation and one-step seedling culture method for single-bud stem segments of pueraria thomsonii | |
| CN103098712B (en) | Davallia mariesii breeding method | |
| CN105028215A (en) | Rooting medium for tissue culture of bletilla striata and tissue culture method of bletilla striata | |
| CN101238793B (en) | A set of culture medium of eucharis grandiflora tissue culture and standardization fast propogation method thereof | |
| CN104396759B (en) | The method that ash tree tissue cultures is bred fast | |
| CN101411305A (en) | Method for quickly reproducing in-vitro pea nut | |
| CN109717065B (en) | Method for promoting germination of alfalfa | |
| CN102067818B (en) | Inducing technology of test tube lotus root | |
| CN111587784A (en) | Rapid propagation method of hydrangea macrophylla | |
| CN1157107C (en) | Fast propagating method of Dendrobium nobile Lindl | |
| CN103348915B (en) | Method for strong seedling and rooting culture of strawberries | |
| CN112400696B (en) | Tissue culture method of evergreen common selfheal fruit-spike bamboo | |
| CN114514879A (en) | Rapid propagation and sugar-free rooting culture method for Nicotiana benthamiana | |
| CN108401905B (en) | Method for rapid propagation culture and annual fresh flower production of gerbera jamesonii | |
| KR20080073388A (en) | Mass production of bulblet via somatic embryogenic cell culture in lily | |
| CN104054579B (en) | A kind of method of tung oil tree petiole directly regenerated plant | |
| CN102754599A (en) | Method for quickly breeding cymbidium hybridium by use of root inducing protocorm | |
| CN103238522B (en) | Open type simplified culture medium for taxus cuspidata | |
| CN111919750A (en) | Tissue culture method of Zanthoxylum piperitum | |
| CN109757378B (en) | Method for rapidly propagating fritillaria thunbergii seedlings by using intermittent immersion system |
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 |