阅读说明：本技术 一种绣球的液体培养基组培快繁方法 (Hydrangea liquid culture medium tissue culture rapid propagation method ) 是由 胡春宏 季翔 常苹 王婷婷 于 2019-11-22 设计创作，主要内容包括：本发明公开了一种绣球的液体培养基组培快繁方法,属于植物组织培养技术领域,旨在提供一种繁殖周期短,效率高,成本低的绣球的液体培养基组培快繁方法,其技术方案要点是具体步骤如下：S1：外植体选择；S2：外植体消毒；S3：诱导培养；S4：增殖培养；S5：生根培养。本发明通过液体组织培养技术,以芽为原材料,获取方便,增殖倍率高达8～10倍,生根率达到95%以上,苗木规格整齐,性状保持稳定,同时节省成本,适合工厂化大规模生产优质绣球种苗。(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.)

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.

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.