CN117694188A - Method for breeding pteris miq spore seedlings - Google Patents

Abstract

The invention relates to a method for breeding golden dog fern spore seedlings, which includes the following steps: S1. Collect mature spores produced by golden dog fern and store them at 4-26°C for later use; S2. Sterilize the substrate Add water to soak it to obtain a germination matrix, which is red jade; S3. After making the suspension of the spores preserved in S1, spray it on the germination matrix described in S2, and culture it under light conditions to obtain Spore seedlings; S4. When the growth of the spore seedlings reaches 1-3 leaves and the plant height is 1-2cm, transplant them into the seedling cultivation medium. The seedling cultivation substrate is peat soil. The seedling cultivation is completed when the roots form tubers. Transfer to understory cultivation. The golden dog fern spores cultured by this method have a high germination rate, and the spore seedlings have a high survival rate when planted under forest.

Description

A method for breeding golden dog fern spore seedlings

Technical field

The present invention relates to the technical field of development and utilization of golden dog fern, and specifically relates to a method for breeding golden dog fern spore seedlings.

Background technique

Goldendoodle fern is a large tree-shaped terrestrial fern of the genus Goldendoodle in the family Pteridaceae. The rhizomes of this plant are used as medicine to nourish the liver and kidneys, strengthen waist and knees, remove rheumatism, strengthen muscles and bones, diuretic and relieve stranguria. The hairs on the stems are Can stop bleeding. The Golden Dog Fern has extremely high ornamental value and medicinal value. It is a national second-level protected fern, and mining is prohibited. Pharmaceutical companies are facing a crisis of shortages and production suspensions. The contradiction between supply and demand of medicinal materials is very prominent. Therefore, it is necessary to study the artificial breeding of Golden Dog Fern. Method is very important.

The reproduction of golden dog fern is greatly affected by season and temperature, and the reproduction coefficient is low, making it difficult to obtain large quantities of seedlings. The spore propagation method is an important method for obtaining large quantities of seedlings, but the spore propagation of Golden Dog Fern has high requirements on environmental conditions. Changes in soil, humidity, light, temperature and other conditions will affect the spore propagation process, and the spores will germinate and emerge less. Troublesome problems such as low gametophyte to sporophyte formation rate, slow development and long nursery time, high proportion of weak seedlings, poor adaptability of seedlings to the natural understory environment, slow growth and even a large number of dead seedlings occur alternately. In the existing technology, CN201910110820.9 provides a method for improving the germination rate of golden retriever spores, but the spores only form prothalli after germination. How to achieve fertilization to form sporophytes in the subsequent key stages, and how to improve the survival rate of sporophyte seedlings. The subsequent issues were not resolved. CN202310738213.3 provides a method for cultivating golden dog fern spores, but does not consider the impact of post-harvest spore storage conditions on the germination rate; and does not consider the difference between the prothallus stage of golden dog fern spores and the growth stage of sporophyte seedlings. Demand limits the speed of seedling cultivation. Therefore, there is an urgent need for a method that can improve the spore germination rate of Golden Dog Fern and the survival rate of spore seedlings under forest planting.

Contents of the invention

The purpose of the present invention is to overcome the existing low germination rate of golden dog fern spores and the low survival rate of spore seedlings planted under forest, and provide a method for breeding golden dog fern spore seedlings.

The specific plans are as follows:

A method for breeding golden dog fern spore seedlings, including the following steps:

S1. Collect the mature spores produced by the golden dog fern and store them at 4-26°C for later use;

S2. Sterilize the substrate and soak it with water to obtain a germination substrate. The germination substrate is red jade;

S3. After making the spores stored in S1 into a suspension, spray it on the germination substrate described in S2, and culture it under light conditions to obtain spore seedlings;

S4. When the growth of the spore seedling reaches 1-3 leaves and the plant height is 1-2cm, transplant it into the seedling cultivation medium. The seedling cultivation substrate is peat soil. When the roots form tubers, the seedlings are completed and transferred to the forest. Cultivation.

Further, the mature spores are stored at 4°C for 0.5-1 year.

Further, the preparation method of the suspension is: sterilize the spores stored in 0.1-0.3gS1, suspend them in an aqueous solution containing an inducer, and shake well.

Further, the inducer is at least one of chlorophyllase, gibberellin, and CaCl ₂ ; the concentration is chlorophyllase 0.2 mg/ml, gibberellin 0.2 mg/ml, and CaCl ₂ 0.4 mg/ml.

Further, the illumination conditions are: the culture temperature is 24-26°C, the illumination is 50-70%, and the illumination time is 10-12h.

Further, the specific method of S4 is: when the growth of the spore seedlings reaches 1-3 leaves and the plant height is 1-2cm, transplant them into the seedling medium to promote rapid growth. Place the spore seedlings in a greenhouse for cultivation every week. Spray water; after 0.5-1 year of cultivation, the roots of the spore seedlings will begin to form tubers and be covered with golden hairs. Cut the seedlings to retain 1-2 leaves, and transfer them to the forest for cultivation together with the non-woven seedling bags.

Further, the initial soil surface cultivated under the forest is covered with mulching film, and a sunshade is built on the seedlings. After 2 to 3 months, the mulch film and sunshade are removed.

Further, the seedling raising substrate is packed into a non-woven seedling raising bag for use.

Further, the conditions for understory cultivation are: selecting fir forests or woodland forests in subtropical areas with annual average temperatures above 5°C.

Beneficial effects:

1. In the present invention, the entire experimental process will not destroy wild resources, and conventional drugs and consumables are used, and the matrix components used are single and easy to obtain;

2. In the present invention, large-scale breeding of spore seedlings through spores produced by Golden Dog Fern under artificial cultivation conditions can greatly improve the spore germination rate and ensure that the prothalli are transformed into sporophytes;

3. The spore seedlings bred by the present invention have a high survival rate when planted under forest.

Description of the drawings

In order to illustrate the technical solution of the present invention more clearly, the accompanying drawings will be briefly introduced below. Obviously, the drawings in the following description only relate to some embodiments of the present invention and do not limit the present invention.

Figure 1 is a growth chart of the underforest culture of the golden retriever fern spore seedlings provided by the present invention;

Figure 2 is a growth chart of the golden dog fern spore seedlings provided by the present invention.

Detailed ways

Preferred embodiments of the invention will be described in more detail below. Although preferred embodiments of the present invention are described below, it should be understood that the present invention may be implemented in various forms and should not be limited to the embodiments set forth herein. If specific techniques or conditions are not specified in the examples, the techniques or conditions described in literature in the field or product instructions will be followed. If the manufacturer of the reagents or instruments used is not indicated, they are all conventional products that can be purchased commercially. In the following examples, "%" refers to weight percentage unless otherwise specified.

This embodiment provides a method for breeding golden dog fern spore seedlings, including the following steps:

S1. Collect the mature spores produced by the golden dog fern and store them at 4-26°C for later use;

S2. Sterilize the substrate and soak it with water to obtain a germination substrate. The germination substrate is red jade;

S3. After making the spores stored in S1 into a suspension, spray it on the germination substrate described in S2, and culture it under light conditions to obtain spore seedlings;

S4. When the growth of the spore seedling reaches 1-3 leaves and the plant height is 1-2cm, transplant it into the seedling cultivation medium. The seedling cultivation substrate is peat soil. When the roots form tubers, the seedlings are completed and transferred to the forest. Cultivation.

In this embodiment, the golden dog fern spores are stored at 4°C for 0.5-1 year. Under this storage condition, the germination rate of the spores can be increased.

In this example, 0.1-0.3g of the golden dog fern spores described in S1 are disinfected and suspended in an aqueous solution containing an inducer, and shaken thoroughly.

In this embodiment, the inducer is at least one of chlorophyllase, gibberellin, and CaCl2; the concentration is chlorophyllase 0.2 mg/ml, gibberellin 0.2 mg/ml, and CaCl ₂ 0.4 mg/ml .

In this embodiment, the illumination conditions are: the culture temperature is 24-26°C, the illumination is 50-70%, and the illumination time is 10-12 hours.

In this embodiment, the specific method of S4 is: when the spore seedlings grow to 1-3 leaves and the plant height is 1-2 cm, they are transplanted into the seedling medium to promote rapid growth, and the spore seedlings are placed in a greenhouse for cultivation. , spray water every week; after 0.5-1 year of cultivation, the roots of the spore seedlings will begin to form tubers and be covered with golden hairs. Cut the seedlings to retain 1-2 leaves, and transfer them to the understory together with the non-woven seedling bags.

In this embodiment, the initial soil surface of the understory culture is covered with mulching film, and a sunshade is built on the seedlings. After 2 to 3 months, the mulch film and sunshade are removed.

In this embodiment, the seedling raising substrate is packed into a non-woven seedling raising bag for use.

In this embodiment, the conditions for understory cultivation are as follows: selecting fir forests or woodland forests in subtropical areas with annual average temperatures above 5°C.

Example 1

Effects of spore germination on golden dog fern stored at different years and temperatures

Use the golden dog fern spores stored under the storage conditions of 4°C for 1 year, 4°C for 2 years, 4°C for 3 years, room temperature for 1 year, and harvested within 48 hours as materials. Take 0.02g of spores and disinfect them and suspend them in 20ml. in distilled water. Use peat soil as the substrate, sow seeds and spray water. Each treatment was repeated three times, and 150 days was used as the observation period to observe the time of prothallus formation and the time of sporophyte emergence. Count when the overall formation rate of prothalli and sporophytes reaches 20 plants. It can be seen from Table 1 that the germination rate of 1-year storage at 4°C is 11 times that of 1-year storage at conventional room temperature, and the germination time is the shortest.

Table 1 Effects of spore germination under storage in different years and temperatures

Example 2

Effects of different substrates on spore germination of Golden Dogfern

For spore germination, peat soil, red jade, vermiculite, and native soil are selected as the substrate. An orthogonal experiment is used. The germination substrate is sterilized in an autoclave pot. After the substrate is cooled, it is put into a culture cup and soaked; take 0.2g, Sterilize the golden dog fern spores stored at 4°C for 1 year and suspend them in 200 ml of distilled water. Sprinkle them evenly on the surface of the substrate, soak them, and place them in an indoor light incubator for culture. The culture temperature is 26°C, the illumination is 66%, and the culture time is 12 hours. . Each treatment was repeated three times, and 150 days was used as the observation period to observe the time of prothallus formation and the time of sporophyte emergence. Count when the overall formation rate of prothalli and sporophytes reaches 20 plants. It can be seen from Table 2 that when red jade is used as the germination medium, the germination rate is 4 times that of the original soil, and the appearance period of prothalli and sporophytes is greatly shortened.

Table 2 Effects of different substrates on golden dog fern spore germination

Example 3

Effects of spores treated with different inducers on germination rate

Prepare respectively 0.4 mg/ml, 20 ml CaCl ₂ solution, 0.2 mg/ml, 20 ml green wood enzyme solution and 0.2 mg/ml, 20 ml gibberellin GA3 solution and 20 ml distilled water. Take 0.2 g of each and store 1 at 4°C. The spores of golden dog fern were suspended in the above four solutions, soaked for 30 minutes, sown in the peat soil matrix, and soaked. Each treatment was repeated 3 times, and 150 days was used as the observation period. The time of formation of prothalli and the appearance of sporophytes were observed. When the overall formation rate of prothalli and sporophytes reached 20 plants, the number was counted. It can be seen from Table 3 that using _CaCl2 solution as an inducer, the spore germination efficiency is significantly improved, which is 2.6 times that of using distilled water and 1.4 times that of using gibberellin GA3 solution. Using green wood enzyme solution as an inducer, the germination rate is 2.2 times that of distilled water and 1.2 times that of gibberellin GA3 solution.

Table 3 Effects of spores treated with different inducers on germination rate

Example 4

Effects of different substrates on the growth of golden dog fern seedlings

In Example 3, the seedlings after germination of golden dog fern spores treated with CaCl ₂ as the inducer were used. When transplanting, peat soil, red jade, vermiculite, and forest soil were used as substrates. Orthogonal experiments were used. Each substrate was filled with non-woven Use a cloth nursery bag, transplant the golden retriever fern spore seedlings into the nursery bag, place it at room temperature for cultivation, water and observe. The growth status of golden dog fern was measured based on the number of leaves, plant height and biomass of the seedlings. It can be seen from Table 4 that when peat soil is used as a seedling cultivation substrate, the plant growth rate is much better than other substrates.

Table 4 Effects of different substrates on the growth of golden retriever seedlings

Example 5

The understory planting is carried out in a fir forest with an annual average temperature above 5°C in the subtropical zone in April. A pit is dug 1m away from the tree trunk, with a pit depth of 15cm and a spacing of 1m between plants. The peat soil used as the substrate in Example 4 is cultivated. Put the golden dog fern seedlings together with the non-woven nursery bag into the pit, backfill the soil and compact it, pour enough water, and cover the soil surface with mulch film. The one without mulch film is used as a control. The mulched group and the non-mulched group are both on the seedlings. Build an awning. After 2 months, remove the mulch and awning. The results showed that after the seedlings went up the mountain, the survival rate of the mulched group was nearly 90% in the first month and nearly 75% after the second month. The survival rate of the non-mulched group was only 25% in the first month and only 10% after the second month. %. In the late survival period, from the 3rd to the 6th month, the survival rate and growth potential of the film-coated group were significantly better than those of the non-coated group.

The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details of the above embodiments. Within the scope of the technical concept of the present invention, a variety of simple modifications can be made to the technical solutions of the present invention. These simple modifications All belong to the protection scope of the present invention.

In addition, it should be noted that the specific technical features described in the above-mentioned specific embodiments can be combined in any suitable manner as long as there is no contradiction. In order to avoid unnecessary repetition, various possible combinations are not further described in the present invention.

In addition, any combination of various embodiments of the present invention can also be carried out. As long as they do not violate the idea of the present invention, they should also be regarded as the disclosed content of the present invention.

Claims (9)

1. A method for breeding pteris british spores is characterized in that: the method comprises the following steps:

s1, collecting mature spores produced by the pteris british, and placing the mature spores at 4-26 ℃ for standby;

s2, soaking the substrate after sterilization treatment by adding water to obtain a germination substrate, wherein the germination substrate is red jade;

s3, preparing the spores stored in the S1 into a suspension, spraying the suspension onto the germination matrix in the S2, and culturing under the illumination condition to obtain spore seedlings;

s4, transplanting the spore seedlings into seedling raising substrates when the growth vigor of the spore seedlings reaches 1-3 leaves and the plant height is 1-2cm, wherein the seedling raising substrates are peat soil, and after roots form tubers, the seedlings are cultivated in a forest.

2. The method for breeding the pteris miq spore seedlings according to claim 1, wherein the method comprises the following steps: the mature spores are stored at 4 ℃ for 0.5-1 year.

3. The method for breeding the pteris miq spore seedlings according to claim 1, wherein the method comprises the following steps: the preparation method of the suspension comprises the following steps: sterilizing 0.1-0.3g of spores stored in S1, suspending in an aqueous solution containing an inducer, and shaking thoroughly.

4. A method for breeding a pteris miq spore seedling according to claim 3, wherein: the saidThe inducer is green xylanase, gibberellin and CaCl ₂ At least one of (a) and (b); the concentration of the chlorazyme is 0.2mg/ml, the gibberellin is 0.2mg/ml and CaCl is 0.2mg/ml ₂ 0.4mg/ml。

5. The method for breeding the pteris miq spore seedlings according to claim 1, wherein the method comprises the following steps: the illumination conditions are as follows: the culture temperature is 24-26 ℃, the illuminance is 50-70%, and the illumination time is 10-12h.

6. The method for breeding the pteris miq spore seedlings according to claim 1, wherein the method comprises the following steps: the specific method of S4 is as follows: transplanting the spore seedlings into a seedling substrate to promote rapid growth when the growth vigor of the spore seedlings reaches 1-3 leaves and the plant height is 1-2cm, placing the spore seedlings into a greenhouse for culture, and spraying water every week; after 0.5-1 year of cultivation, the root of the spore seedling starts to form tubers, golden villus is covered, the seedling is sheared to keep 1-2 leaves, and the seedlings are transferred into the under-forest cultivation together with a non-woven fabric seedling bag.

7. The method for breeding the pteris miq spore seedlings according to claim 6, wherein the method comprises the following steps: and covering the initial soil surface of the under-forest culture with a mulching film, building a sunshade on the seedlings, and removing the mulching film and the sunshade after 2-3 months.

8. The method for breeding the pteris miq spore seedlings according to claim 6, wherein the method comprises the following steps: the seedling substrate is put into a non-woven fabric seedling bag for use.

9. The method for breeding the pteris miq spore seedlings according to claim 6, wherein the method comprises the following steps: the conditions of the culture under the forest are as follows: selecting fir forest or wood lotus forest in subtropical region with average annual temperature above 5 deg.C.