CN110839529A - Hydrangea liquid culture medium tissue culture rapid propagation method - Google Patents
Hydrangea liquid culture medium tissue culture rapid propagation method Download PDFInfo
- Publication number
- CN110839529A CN110839529A CN201911161718.8A CN201911161718A CN110839529A CN 110839529 A CN110839529 A CN 110839529A CN 201911161718 A CN201911161718 A CN 201911161718A CN 110839529 A CN110839529 A CN 110839529A
- Authority
- CN
- China
- Prior art keywords
- culture
- hydrangea
- medium
- tissue culture
- explant
- 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.)
- Pending
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
- A01H4/001—Culture apparatus for tissue culture
Abstract
The invention discloses a liquid culture medium tissue culture rapid propagation method of hydrangea, belongs to the technical field of plant tissue culture, and aims to provide a liquid culture medium tissue culture rapid propagation method of hydrangea with short propagation period, high efficiency and low cost, wherein the technical scheme is characterized by comprising the following steps: s1: selecting an explant; s2: sterilizing explants; s3: performing induction culture; s4: carrying out proliferation culture; s5: and (5) rooting culture. According to the method, the liquid tissue culture technology is adopted, the buds are used as raw materials, the method is convenient to obtain, the multiplication rate is up to 8-10 times, the rooting rate reaches more than 95%, the seedling size is neat, the character is stable, the cost is saved, and the method is suitable for industrial large-scale production of high-quality hydrangea seedlings.
Description
Technical Field
The invention relates to the technical field of plant tissue culture, in particular to a hydrangea liquid culture medium tissue culture rapid propagation method.
Background
Hydrangea (Hydrangea macrophylla) is deciduous shrub of Hydrangea of Saxifragaceae, and is called Hydrangea, Astrongylus, Maackia sida, snowball, Hydrangea, and the like, and is an ornamental plant widely applied to gardens. Native to the Yangtze river basin and south of China, and the natural plant height is 2 m. Leaf pair, inverted egg or ellipse, sawtooth at edge, 20cm diameter, and nearly spherical. The flower color is changeable, bluish white, gradually turns pink, and then turns purple red, and the flower color is beautiful and gorgeous. The color of the soil-base composite material is changed along with the pH value of the soil, and when the pH value of the soil is 4-6, the color of the soil-base composite material is mostly blue; when the pH value is above 7.5, the color is red. The hydrangea leaves are verdure, the flower color is bright, the flowers are in a cluster and beautiful and colorful when the flowers are full, each cluster of flowers can be opened for 2 months, and the ornamental period is long.
The pot-growing flowers of hydrangea are few and are not easy to seed, and are usually propagated by a plant division, layering or cuttage method, but the propagation rate of the plant division and layering is low, the speed is slow, the cuttage propagation is limited by seasons, seedlings cannot be produced all the year round, and the market demand is difficult to meet.
Disclosure of Invention
Aiming at the defects in the prior art, the invention aims to provide a hydrangea liquid culture medium tissue culture rapid propagation method with short propagation period, high efficiency and low cost.
The above object of the present invention is achieved by the following technical solutions:
a liquid culture medium tissue culture rapid propagation method of hydrangea, comprising the following steps:
s1: selecting an explant;
s2: sterilizing explants;
s3: performing induction culture;
s4: carrying out proliferation culture;
s5: rooting and culturing to obtain the root-growing culture medium,
wherein, in S3, the explant sterilized in S2 is inoculated into an induction medium for culture, and the formula of the induction medium is MS +6-BA 0.5mg/L + GA32.0mg/L + sucrose 30g/L, the pH value of the induction culture medium is 5.80,
in S4, the sterile explant in S3 is transferred to a proliferation medium, the formula of the proliferation medium is MS +6-BA 0.3-1.0 mg/L + GA 30.5-1.5 mg/L + sucrose 30g/L, the pH value of the proliferation medium is 5.80,
in S5, the hydrangea tissue culture seedling in S4 is placed into a rooting medium, the formula of the rooting medium is 1/2MS + IBA 0.5-2.0 mg/L + NAA 1.0-2.0 mg/L + sucrose 15g/L, and the pH value of the rooting medium is 5.80.
By adopting the technical scheme, the buds are used as raw materials through a liquid tissue culture technology, the method is convenient to obtain, the multiplication rate is up to 8-10 times, the rooting rate reaches more than 95%, the nursery stock is neat in specification, the characters are kept stable, the cost is saved, and the method is suitable for industrial large-scale production of high-quality hydrangea seedlings. The germination rate of the induction culture medium can reach 90 percent, and a sterile regeneration system can be successfully established. By using the multiplication culture medium, the multiplication rate of hydrangea tissue culture seedlings can reach 8-10 times, the height of the tissue culture seedlings is consistent, and the tissue culture seedlings grow robustly. By using the rooting culture medium, the rooting rate of the hydrangea tissue culture seedlings can reach 95%, the root system is developed and robust, and the hydrangea tissue culture seedlings can be successfully used for transplanting greenhouses.
The present invention in a preferred example may be further configured to: in S3, the culture environment after the explant disinfected in S2 is inoculated into an induction culture medium is under the condition of the illumination intensity of 1500Lux, the temperature is 25 ℃, and the illumination time is 16 h; culturing for 6-8 weeks under the dark condition at 23 ℃ under the illumination time of 8 h.
By adopting the technical scheme, the stress reaction of the plant material in the induction culture stage can be effectively avoided under the culture condition, the plant material can be quickly adapted to an induction culture medium, and the plant material is promoted to quickly grow and the bud is strong.
The present invention in a preferred example may be further configured to: in S4, the culture environment after inoculating the sterile explant in S3 into the proliferation medium is under the condition of the illumination intensity of 1500Lux, the temperature is 25 ℃, and the illumination time is 16 h; culturing for 8-12 weeks under the dark condition at 23 ℃ under the illumination time of 8 h.
By adopting the technical scheme, the plant material can be effectively prevented from generating stress reaction in the propagation culture stage under the culture condition, and the plant material can be quickly adapted to a propagation culture medium, so that the plant material can quickly grow, and the bud bodies are strong.
The present invention in a preferred example may be further configured to: in S4, a new growth medium is inoculated every 8 to 12 weeks.
By adopting the technical scheme, after 8-12 weeks, the effect of the proliferation culture medium is weakened, and plant materials can pollute the proliferation culture medium and need to be replaced in time.
The present invention in a preferred example may be further configured to: in S5, the culture environment after the hydrangea tissue culture seedlings in S4 are inoculated into the rooting culture medium is under the condition of illumination intensity of 1500Lux, the temperature is 25 ℃, and the illumination time is 16 h; culturing for 8-12 weeks under the dark condition at 23 ℃ under the illumination time of 8 h.
By adopting the technical scheme, the stress reaction of the plant material in the rooting culture stage can be effectively avoided under the culture condition, the plant material can be quickly adapted to the rooting culture medium, and the plant material is promoted to quickly grow and the bud is strong.
The present invention in a preferred example may be further configured to: in S2, the specific steps of explant disinfection are that leaves of raspberry explants are cut off, the leaves are washed clean with tap water, the washed clean explants are immersed in 75% alcohol on a workbench for disinfection for 30S, the explants are washed with sterile water for 3-4 times, white cat bleaching water and sterile water are used according to the ratio of 1: 4, preparing a disinfectant, and soaking the explant in the disinfectant for 40 min.
By adopting the technical scheme, the method for disinfecting the explant is simple, the death rate of the explant can be greatly reduced, and the disinfection survival rate is high.
The present invention in a preferred example may be further configured to: in S1, the explant is selected to be the stem tip portion of a semi-lignified shoot with a plump shoot bud of a healthy hydrangea.
In summary, the invention includes at least one of the following beneficial technical effects:
according to the method, the liquid tissue culture technology is adopted, the buds are used as raw materials, the method is convenient to obtain, the multiplication rate is up to 8-10 times, the rooting rate reaches more than 95%, the seedling size is neat, the character is stable, the cost is saved, and the method is suitable for industrial large-scale production of high-quality hydrangea seedlings.
Drawings
FIG. 1 is a schematic flow chart of a liquid medium tissue culture rapid propagation method of hydrangea.
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings.
Example (b): referring to fig. 1, the method for tissue culture and rapid propagation of hydrangea by using a liquid culture medium disclosed by the invention comprises the following specific steps:
(I), test materials: the test material was collected from healthy hydrangea in nursery garden of Jiangsu Dongyu plant science and technology Limited, the variety was Endless summer (bud struck), and the explant was half lignified shoot tip with plump current-year shoot bud.
(II) test method:
the hydrangea tissue culture process is carried out according to the following steps: primary culture, secondary culture and rooting culture.
(1) And primary culture: the hydrangea explant is cut off, and washed clean with tap water. And (3) immersing the washed explants on a clean bench into 75% alcohol for disinfection for 30s, and washing with sterile water for 3-4 times. White cat bleach water and sterile water were used at a ratio of 1: 4, soaking the explant in the disinfectant for 40min, wherein the white cat bleaching water is 'white cat' brand domestic bleaching water produced by Shanghai white cat (group) limited company. After the disinfection is finished, inoculating the explant into an induction culture medium for culture, and establishing a sterile regeneration system, wherein the induction culture environment is under the condition of illumination intensity of 1500Lx, the temperature is 25 ℃, and the illumination time is 16 h; culturing for 6-8 weeks under the dark condition at 23 ℃ under the illumination time of 8 h. Wherein the induction culture medium is MS +6-BA 0.5-1.5 mg/L + GA31.0-3.5 mg/L + sucrose 30g/L, and the pH value of the induction culture medium is 5.80.
(2) And subculturing: taking the established sterile regeneration system as an object, transferring the sterile explant to a multiplication medium, wherein the multiplication medium is MS +6-BA 0.3-1.0 mg/L + GA 30.5-1.5 mg/L + sucrose 30g/L, and the culture environment is under the condition of illumination intensity of 1500Lx, the temperature is 25 ℃, and the illumination time is 16 h; culturing for 8-12 weeks under the dark condition at 23 ℃ under the illumination time of 8 h. And (3) inoculating a new proliferation culture medium every 8-12 weeks, wherein the pH value of the proliferation culture medium is 5.80.
(3) And (3) rooting culture: taking terminal buds of 2-3 cm of hydrangea tissue culture seedlings subjected to subculture, and putting the terminal buds into a rooting culture medium, wherein the rooting culture medium is 1/2MS, IBA is 0.5-2.0 mg/L, NAA is 1.0-2.0 mg/L, sucrose is 15g/L, and the pH value of the rooting culture medium is 5.80. The induction culture environment is under the condition of illumination intensity of 1500Lx, the temperature is 25 ℃, and the illumination time is 16 h; culturing for 8-12 weeks under the dark condition at 23 ℃ under the illumination time of 8 h.
(III) test results:
primary culture: by using the disinfection method, the disinfection success rate can reach 80%, the germination rate of the induction culture medium can reach 90%, and a sterile regeneration system can be successfully established.
Subculturing: by using the multiplication medium disclosed by the invention, the multiplication rate of hydrangea tissue culture seedlings can reach 8-10 times, the height of the tissue culture seedlings is consistent, and the tissue culture seedlings grow robustly.
Rooting culture: by using the rooting culture medium, the rooting rate of the hydrangea tissue culture seedlings can reach 95%, the root system is developed and robust, and the rooting culture medium can be successfully used for transplanting greenhouses.
The MS culture medium of the invention is a culture medium developed by Murashige and Skoog, and the composition formula of the MS culture medium is shown in Table 1.
1/2MS culture medium contains macroelements half of MS culture medium, and other components are used in the same amount.
TABLE 1 ingredient Table of MS culture Medium
The plant growth regulator related to the invention comprises: 6-BA-6-benzylaminopurine; GA3-gibberellin; IBA-indolebutyric acid, NAA-naphthylacetic acid.
The induction culture medium, the multiplication culture medium and the rooting culture medium are all liquid culture media.
The embodiments of the present invention are preferred embodiments of the present invention, and the scope of the present invention is not limited by these embodiments, so: all equivalent changes made according to the structure, shape and principle of the invention are covered by the protection scope of the invention.
Claims (7)
1. A hydrangea liquid culture medium tissue culture rapid propagation method is characterized in that: the method comprises the following specific steps:
s1: selecting an explant;
s2: sterilizing explants;
s3: performing induction culture;
s4: carrying out proliferation culture;
s5: rooting and culturing to obtain the root-growing culture medium,
wherein in S3, the explant disinfected in S2 is inoculated into an induction culture medium for culture, and the formula of the induction culture medium is MS +6-BA 0.5-1.5 mg/L + GA31.0-3.5 mg/L + sucrose 30g/L, the pH value of the induction culture medium is 5.80,
in S4, the sterile explant in S3 is transferred to a proliferation medium, the formula of the proliferation medium is MS +6-BA 0.3-1.0 mg/L + GA 30.5-1.5 mg/L + sucrose 30g/L, the pH value of the proliferation medium is 5.80,
in S5, the hydrangea tissue culture seedling in S4 is placed into a rooting medium, the formula of the rooting medium is 1/2MS + IBA 0.5-2.0 mg/L + NAA 1.0-2.0 mg/L + sucrose 15g/L, and the pH value of the rooting medium is 5.80.
2. The method for rapid propagation of hydrangea by tissue culture in liquid medium according to claim 1, wherein: in S3, the culture environment after the explant disinfected in S2 is inoculated into an induction culture medium is under the condition of the illumination intensity of 1500Lux, the temperature is 25 ℃, and the illumination time is 16 h; culturing for 6-8 weeks under the dark condition at 23 ℃ under the illumination time of 8 h.
3. The method for rapid propagation of hydrangea by tissue culture in liquid medium according to claim 1, wherein: in S4, the culture environment after inoculating the sterile explant in S3 into the proliferation medium is under the condition of the illumination intensity of 1500Lux, the temperature is 25 ℃, and the illumination time is 16 h; culturing for 8-12 weeks under the dark condition at 23 ℃ under the illumination time of 8 h.
4. The method for rapid propagation of hydrangea by tissue culture in liquid medium according to claim 3, wherein: in S4, a new growth medium is inoculated every 8 to 12 weeks.
5. The method for rapid propagation of hydrangea by tissue culture in liquid medium according to claim 1, wherein: in S5, the culture environment after the hydrangea tissue culture seedlings in S4 are inoculated into the rooting culture medium is under the condition of illumination intensity of 1500Lux, the temperature is 25 ℃, and the illumination time is 16 h; culturing for 8-12 weeks under the dark condition at 23 ℃ under the illumination time of 8 h.
6. The method for rapid propagation of hydrangea by tissue culture in liquid medium according to claim 1, wherein: in S2, the specific steps of explant disinfection are that leaves of raspberry explants are cut off, the leaves are washed clean with tap water, the washed clean explants are immersed in 75% alcohol on a workbench for disinfection for 30S, the explants are washed with sterile water for 3-4 times, white cat bleaching water and sterile water are used according to the ratio of 1: 4, preparing a disinfectant, and soaking the explant in the disinfectant for 40 min.
7. The method for rapid propagation of hydrangea by tissue culture in liquid medium according to claim 1, wherein: in S1, the explant is selected to be the stem tip portion of a semi-lignified shoot with a plump shoot bud of a healthy hydrangea.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911161718.8A CN110839529A (en) | 2019-11-22 | 2019-11-22 | Hydrangea liquid culture medium tissue culture rapid propagation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911161718.8A CN110839529A (en) | 2019-11-22 | 2019-11-22 | Hydrangea liquid culture medium tissue culture rapid propagation method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110839529A true CN110839529A (en) | 2020-02-28 |
Family
ID=69604050
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911161718.8A Pending CN110839529A (en) | 2019-11-22 | 2019-11-22 | Hydrangea liquid culture medium tissue culture rapid propagation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110839529A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112369315A (en) * | 2020-11-17 | 2021-02-19 | 中南林业科技大学 | Breeding method of hydrangea |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0325816A1 (en) * | 1988-01-05 | 1989-08-02 | Leo Anne Eveleens | Method for forcing hydrangea plants into bloom all year round |
| CN106212286A (en) * | 2016-08-10 | 2016-12-14 | 江苏绿洲园艺绿化有限公司 | A kind of laurustinus special culture media |
| CN109618925A (en) * | 2018-11-07 | 2019-04-16 | 中南林业科技大学 | A method of suitable for a variety of shrubs and herbage flower micropropagation of plants |
-
2019
- 2019-11-22 CN CN201911161718.8A patent/CN110839529A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0325816A1 (en) * | 1988-01-05 | 1989-08-02 | Leo Anne Eveleens | Method for forcing hydrangea plants into bloom all year round |
| CN106212286A (en) * | 2016-08-10 | 2016-12-14 | 江苏绿洲园艺绿化有限公司 | A kind of laurustinus special culture media |
| CN109618925A (en) * | 2018-11-07 | 2019-04-16 | 中南林业科技大学 | A method of suitable for a variety of shrubs and herbage flower micropropagation of plants |
Non-Patent Citations (2)
| Title |
|---|
| 李杨丽: "八仙花离体快繁及盆栽生产相关技术研究", 《中国硕士学位论文全文数据库农业科技辑》 * |
| 王忠武、建德锋: "八仙花茎尖离体培养技术研究", 《北方园艺》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112369315A (en) * | 2020-11-17 | 2021-02-19 | 中南林业科技大学 | Breeding method of hydrangea |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104012417B (en) | High-efficiency and rapid micropropagation method for toxicodendron vernicifluum | |
| CN106417015B (en) | A kind of Huaiji primulina tabacum tissue cultures and rapid propagation method | |
| CN110521600B (en) | Method for establishing efficient sterile regeneration system by using curcuma alismatifolia stalks | |
| CN101822220A (en) | Method for culturing and rapidly propagating stem tip tissue of rare cymbidium goeringii | |
| CN103430842B (en) | A kind of Quick propagation method of hybrid orchid tissue culture | |
| CN108029559B (en) | Method for quickly cultivating ilex latifolia tissue culture seedlings | |
| CN106857256A (en) | The method that beautiful dew breeding potential is improved based on callus induction Regeneration Ways | |
| CN108703073A (en) | Southerm yew tissue cultivating and seedling method | |
| CN102210267A (en) | Method for regenerating rose into complete plant | |
| CN103733995A (en) | Peony callus induction method | |
| CN110583488A (en) | Method for establishing tissue culture rapid propagation technical system of new lycoris variety' pink | |
| CN102415339A (en) | Rapid propagation method of photinia fraseri | |
| CN103651141A (en) | Factory-like rapid propagation method for test-tube seedlings of dendranthema morifolium | |
| CN110810241A (en) | Tissue culture seedling propagation method for raspberries | |
| CN110839529A (en) | Hydrangea liquid culture medium tissue culture rapid propagation method | |
| CN109220809B (en) | Koelreuteria paniculata somatic embryogenesis and plant regeneration culture method | |
| CN101836589B (en) | Method of rapid propagation of populus | |
| CN111587784A (en) | Rapid propagation method of hydrangea macrophylla | |
| CN104396756B (en) | A kind of fox-brush orchid bud point quick breeding method for tissue culture | |
| CN117158315A (en) | Tissue culture method of gardenia lobule | |
| CN111134019A (en) | Method for directly generating somatic embryos and regenerating plants of ilex denticulata | |
| CN108271687B (en) | Tissue culture method of cerasus serrulata | |
| CN111165354B (en) | Tissue culture propagation method of xanthoceras sorbifolia bunge | |
| CN114027182A (en) | Tissue culture propagation method for dolichos succulent plants in crassulaceae echeveria | |
| CN108064697B (en) | Efficient induction and transplanting method for aralia elata tissue culture seedlings |
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 | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200228 |