CN113767848A

CN113767848A - Efficient rooting method for rhododendron lapponicum tissue culture seedlings

Info

Publication number: CN113767848A
Application number: CN202110982306.1A
Authority: CN
Inventors: 朱娇; 吕晓惠; 董飞; 王烨楠; 齐宇; 王成鹏; 温超; 马蕾
Original assignee: Shandong Academy of Agricultural Sciences
Current assignee: Shandong Academy of Agricultural Sciences
Priority date: 2021-08-25
Filing date: 2021-08-25
Publication date: 2021-12-10

Abstract

The invention discloses a method for efficiently rooting tissue culture seedlings of alpine rhododendrons, which is characterized in that the tissue culture seedlings of alpine rhododendrons are cultivated by utilizing a pure water moss substrate planting technology, so that the rooting efficiency of the tissue culture seedlings is improved to 90%, the number of lateral roots is obviously improved, the survival rate is improved to 95%, the rooting time is shortened, the operation is convenient, and the industrialized large-area popularization is facilitated.

Description

Efficient rooting method for rhododendron lapponicum tissue culture seedlings

Technical Field

The invention relates to a method for efficiently rooting tissue culture seedlings of alpine rhododendrons, and belongs to the technical field of plant tissue culture.

Background

Rhododendron Delavayi (Rhododendron Delavayi) refers to the evergreen Rhododendron in the subfamily of anaphalis, Rhododendron and Rhododendron in the subfamily of Rhododendron and new horticultural variety cultivated by hybridization of the three subfamilies for hundreds of years. The method is originally produced in the alpine forest in the areas of the middle and the west of China, then introduced by Germany, Belgium and other countries for cultivation and domestication, and improves and breeds a plurality of excellent varieties, so the ornamental value is extremely high, and the method is very attractive in the flower market at home and abroad in recent years. The alpine rhododendron usually adopts cuttage and grafting propagation, but many rare varieties are difficult to root and the grafting is difficult to survive, and the alpine rhododendron is not suitable for mass commercial production due to the limitation of seasons, parent plant materials and the like. The alpine rhododendron sold in the market at present mainly depends on import, and the cost of imported finished flowers is high, so that the price of the alpine rhododendron is high.

With the development of plant tissue culture technology, the tissue culture of alpine rhododendron is also widely concerned by researchers, and a great deal of research and progress is made on the tissue culture of rhododendron at home and abroad. However, tissue culture of alpine rhododendron usually has a series of problems that explants are easy to brown, have high vitrification rate, are difficult to root and the like. Therefore, the technical problem of difficult efficient rooting is urgently needed to be broken through. The rooting rate of rhododendron lapponicum is improved, so that technical support is provided for large-scale industrialization of the rhododendron lapponicum.

Disclosure of Invention

The invention overcomes the defects of the prior art, provides a method for efficiently rooting tissue culture seedlings of alpine rhododendron, utilizes a pure water moss substrate planting technology to cultivate the tissue culture seedlings of alpine rhododendron, improves the rooting efficiency of the tissue culture seedlings to 90 percent, obviously improves the number of lateral roots, improves the survival rate to 95 percent, shortens the rooting time, is convenient to operate, and is beneficial to industrial large-area popularization.

A method for efficiently rooting tissue culture seedlings of rhododendron lapponicum comprises the following steps:

1) selecting tissue culture cluster buds of alpine rhododendron, cutting bud seedlings, inoculating the bud seedlings into a buffering rooting culture medium, carrying out dark culture for 5-7d, putting the bud seedlings into a greenhouse, covering non-woven fabrics, and hardening the seedlings for 2-3 d;

2) sterilizing the bud seedlings treated in the step 1) and then airing;

3) soaking the sphagna substrate with deionized water for 3-4 hours, drying to remove water to obtain a soaked sphagna substrate, wrapping the base of the sprout with the soaked sphagna substrate, transplanting the sprout into a plug tray, covering with a plastic film, spraying water for 3-4 days, spraying water for 13-15 days, spraying water for each week, culturing for 40-42 days, and growing into a seedling;

4) transplanting the seedling into the culture medium for conventional culture when the seedling grows to 10-12cm in height and the root system grows to 3-5 cm.

Further, the bud seedling in the step 1) is about 3-5cm long and has 4-5 leaves.

Further, the buffered rooting culture medium in the step 1) is prepared by adding the following components based on a WPM minimal medium: NAA IBA 0.25mg/L:0.25mg/L, 30g/L of soft sugar, 1g/L of activated carbon and 6g/L of agar powder.

Further, the temperature in the greenhouse in step 1) is kept at 25-28 ℃.

Further, the sterilization in step 2) is: soaking the bud seedling in 800 times diluted carbendazim disinfectant for 12-15 min.

Further, the water throwing-off in the step 3) is that a small amount of water overflows until the holding is tight.

Further, the culture medium in the step 4) is formed by mixing peat and perlite according to the mass ratio of (0.8-1) to (0.8-1).

Has the advantages that:

according to the application, the rhododendron lapponicum tissue culture seedlings are cultivated by using a pure water moss substrate planting technology, the rooting rate of the rhododendron lapponicum tissue culture seedlings reaches 90%, the main root systems and the lateral root branches of the tissue culture seedlings obtained by the method are large in number, and the survival rate is improved to 95%.

Drawings

FIG. 1 is a diagram of sterile tissue culture of roselle buds.

FIG. 2 shows the rooting state of alpine rhododendron on pure water moss substrate.

FIG. 3 shows the comparison of the root system of pure water moss planting with a small amount of water moss wrapped and peat wrapped.

FIG. 4 g4 shows the results of processing of alpine rhododendron in the presence of pure sphagna, peat and peat: and (3) rooting of perlite on three substrates of 2: 1.

Detailed Description

In order to make the technical solutions in the present application better understood, the present invention is further described below with reference to examples, which are only a part of examples of the present application, but not all examples, and the present invention is not limited by the following examples.

Example 1 planting technique of pure water moss matrix and planting technique of existing matrix containing a small amount of water moss

1. Experimental procedure

1) Material and treatment: selecting healthy and pollution-free rhododendron delavayi tissue culture cluster buds (as shown in figure 1), cutting a bud seedling with 4-5 leaves of about 3-5cm on a super-clean workbench by using a scalpel, and inoculating the bud seedling into a buffering rooting culture medium, wherein the buffering culture medium is as follows: the WPM basic culture medium is added with NAA (American agar-agar), IBA (0.25 mg/L) and 0.25mg/L, soft sugar 30g/L, active carbon 1g/L and agar powder 6 g/L. Dark culture is carried out for 7 days;

2) placing the tissue culture bottle in an intelligent greenhouse at the temperature of 25-28 ℃, covering the tissue culture bottle with white non-woven fabrics, hardening the seedlings for 3d, taking out the bud seedlings, soaking the bud seedlings for 15min by 800 times of carbendazim disinfectant, placing the bud seedlings on newspaper, and airing the bud seedlings for later use;

3) the pure water moss matrix planting technology comprises the following steps: (1) treating the sphagna: soaking the sphagna substrate with deionized water (EC ═ 0) for 4 hours, then spin-drying for 2 minutes by using a centrifuge, and preferably, holding the sphagna substrate tightly by hands until a little water overflows; (2) wrapping sprout base 1.5cm with soaked pure water moss matrix, transplanting into white transparent 50-hole plug (plug specification: upper caliber: 4.8 × 4.8, lower caliber: 2.3 × 2.3, depth: 9cm, gram weight: 150g), compactness 80%, covering with plastic film, spraying deionized water (EC: 0) every 3 days, spraying once every 2 weeks, culturing for 40 days, randomly digging out 10 plants, observing and counting rooting rate, and counting rooting rate

4) A small amount of sphagna matrix wrapping and peat wrapping planting technology: (1) treating the sphagna: soaking the sphagna substrate with deionized water (EC ═ 0) for 4 hours, then spin-drying for 2 minutes by using a centrifuge, and preferably, holding the sphagna substrate tightly by hands until a little water overflows; (2) wrapping the base part of the bud seedling by using a soaked pure water moss matrix for 1.5cm, wherein the weight of the water moss matrix is about 10g, wrapping the peat matrix for about 120g, transplanting the soaked pure water moss matrix into a white transparent plug with 50 holes (the plug specification is that the upper caliber is 4.8 multiplied by 4.8, the lower caliber is 2.3 multiplied by 2.3, the depth is 9cm, the gram weight is 150g), the compactness is 80%, covering a plastic film, spraying deionized water (EC 0) every 3 days, spraying once every 1 week after culturing for 40 days, randomly digging 10 plants out, observing and counting the rooting condition and the number of roots after culturing for 40 days;

5) transplanting the seedlings to peat when the height of the tissue culture seedlings reaches 10-12cm and the root system grows to 3-5 cm: the perlite is cultured in a matrix of 1:1 in a conventional way.

2. Results of the experiment

Comparing the rooting rate and root system condition of alpine rhododendron after pure water moss planting and small amount of water moss wrapping and peat wrapping planting, as shown in the left side of fig. 3, the rooting rate of alpine rhododendron is 90% -100% for pure water moss wrapping and small amount of alpine rhododendron is 80% -85% for water moss wrapping and peat wrapping planting, the root system quantity of pure water moss wrapping is about 50-60, the root system length is 5-6cm, the lateral root and fibrous root quantity is large, the root system quantity of small amount of water moss wrapping and peat wrapping planting is about 10-20, the root system length is 0.5-1.5cm, the lateral root and fibrous root quantity is small, and the root system quantity is small. Therefore, the pure water moss wrapped planting is superior to a small amount of water moss wrapped and peat wrapped planting in terms of rooting rate or root system state.

In addition, the amount of the water moss planted by the pure water moss matrix is large, the weight of a single cup is about 150g, the wrapping amount of the water moss part is about 10g, the water moss has higher water absorption and water retention, the watering frequency in later management is less, the water moss and peat matrix are fewer, the physicochemical property of the water moss matrix is more stable, loose and breathable, and the growth of the root system of the rhododendron lapponicum tissue culture seedling is more facilitated.

Example 2

1. Materials: healthy pollution-free alpine rhododendron tissue culture cluster buds

2. Method of producing a composite material

(1) The treatment method of the rhododendron lapponicum cluster buds comprises the following steps: cutting a bud seedling with 4-5 leaves of about 3-5cm on a super clean bench by using a scalpel, and inoculating the bud seedling to a buffer rooting minimal medium, wherein the culture medium comprises the following components in parts by weight: WPM basic culture medium, adding soft white sugar 30g/L, activated carbon 1g/L, agar powder 6g/L, and simultaneously adding 4 (g1, g2, g3, g4) buffered rooting culture media with different hormone ratios, wherein (g1 ═ NAA: IBA ═ 1.5:0.5), g2 ═ NAA: IBA ═ 1.0:0.5), g3 ═ NAA: IBA ═ 0.5:0.5), g4 (NAA: IBA ═ 0.25:0.25)) are placed in each bottle, each culture medium is inoculated into 5 bottles, repeated three times, and then the treatment is placed in a tissue culture room under dark conditions, and the treatment is carried out for 7d, and the culture temperature is 25-28 ℃.

(2)7d, placing the tissue culture bottle in an intelligent greenhouse, covering the tissue culture bottle with white non-woven fabrics at the temperature of 25-30 ℃, hardening the seedlings for 3d, taking out the seedlings, soaking the seedlings for 15min by using 800 times of carbendazim disinfectant, placing the soaked seedlings on newspaper, airing the seedlings, transplanting the seedlings into three different matrixes of 2:1, namely pure water moss, peat and perlite, planting the seedlings in a white transparent plug tray with 50 holes, covering a plastic film, and keeping the humidity to be higher than 90% and the temperature to be 25-30 ℃. Deionized water (EC ═ 0) was sprayed every 3 days, and after 2 weeks, deionized water (EC ═ 0) was sprayed every week, and the rooting rate was counted after 40 days of culture.

3) After the root system of the tissue culture seedling grows to 3-5cm, transplanting the seedling into a matrix of peat and perlite which are 1:1 for conventional culture.

3. Conclusion

The method uses pure water moss for wrapping and planting, the rooting rate of the tissue culture seedling of the alpine rhododendron reaches more than 90%, wherein the g4 treatment is the best and is 100%, the main root system and the lateral root branches of the tissue culture seedling obtained by the treatment have a large number, and the survival rate is improved to 95%. (FIG. 4)

TABLE 1 rooting rate, survival rate and plant status of Rhododendron lapponicum tissue culture seedlings treated with different hormones in different substrates

Claims

1. A method for efficiently rooting tissue culture seedlings of alpine rhododendron is characterized by comprising the following steps:

2) sterilizing the bud seedlings treated in the step 1) and then airing;

2. A method as claimed in claim 1 wherein the sprouts of step 1) are about 3-5cm long with 4-5 leaves.

3. The method of claim 1, wherein the buffered rooting medium of step 1) is based on WPM minimal medium supplemented with the following components: NAA IBA 0.2-0.25 mg/L: 0.2-0.25mg/L, 25-30g/L of soft sugar, 0.8-1g/L of activated carbon and 5-6g/L of agar powder.

4. The method as claimed in claim 1, characterized in that the temperature in the greenhouse in step 1) is kept between 25 and 28 ℃.

5. The method as set forth in claim 1, wherein the sterilization in the step 2) is: soaking the bud seedling in 800 times diluted carbendazim disinfectant for 12-15 min.

6. The method of claim 1 wherein the spin-dry moisture in step 3) is such that a small amount of water spills out of the grip.

7. The method as claimed in any one of claims 1 to 6, wherein the cultivation substrate in step 4) is peat and perlite mixed in a mass ratio of (0.8-1) to (0.8-1).