CN105961010B - Utilize the method for orchid root bacteria cultivation hectolitre pine - Google Patents

Abstract

The invention provides a method for cultivating male stone pine with orchid rhizomes. Clean the orchid mycorrhizae with tap water, sterilize the surface with 75% alcohol, cut out the nodular tissue on the ultra-clean workbench, and move it into the fungus plate culture medium to wait for mycelia Grow out, extract mycelia and transfer to the same plate medium for purification and culture; culture at 25-30°C for 7-8 days to cover the plate medium, cut the mycelium block on the plate and inoculate it into the mixed bacteria culture bag; Cultivate at 28°C for 35‑40 days until the mycelium grows through the bacteria bag; mix the mycelium through the bacteria bag with the base material planted with orchids, and use it as the male stone pine cultivation substrate; then inoculate the male stone pine tissue culture seedlings into the substrate To cultivate. The present invention uses the seedlings of wild male stone pine through tissue culture, and adopts planted orchid base material to add artificially cultivated endophytic fungus mixed substrate to cultivate male stone pine, which can obviously promote the increase of plant height and weight and improve product quality, so as to better develop and utilize The medicinal value of male stone pine.

Description

The method of cultivating male stone pine with orchid root fungus

technical field

The invention belongs to the field of plant cultivation, and in particular relates to a method for cultivating male stone pine by using orchid root fungus.

Background technique

Gongshisong: Ludisia discolor ( Ker-Gawl. ) A. Rich., a plant of Orchidaceae, is a herbal medicine with local characteristics in southern Fujian. It has the effects of clearing heat and cooling blood, promoting body fluid and reducing fire, diuresis and collaterals, and relieving cough. It can be used to treat persistent high fever, sore throat, swelling and pain without name, spleen and stomach diseases and inflammation. "Chinese Materia Medica" records its efficacy: "moisten the lungs, invigorate the spleen, calm the nerves, and treat mainly tuberculosis and hemoptysis, neurasthenia, and loss of appetite." Male stone pine has strict requirements on the growth environment, and often grows in damp rock crevices, and the growth place is mostly rich in endophytic fungi. Its growth process requires the formation of a symbiotic relationship with fungi.

Artificial cultivation According to some dealers' investigations on the artificial cultivation of fresh plants of male stone pine, because the male stone pine has strict requirements on the growth environment, there is no more authoritative mature technology to refer to, and few people have carried out cultivation and trial planting research. Planting Techniques. The male stone pine sold in the market is generally cultivated in three forms of sand culture, hydroponics and soil culture for short-term cultivation by purchasing fresh plants. Sand culture is used by most people because of its good air permeability, sufficient mineral nutrition, less water accumulation and no root rot, etc., but the general technical content is low, and the growth of biological yield is not obvious. Moreover, sand culture is not used for large-scale seedling cultivation, but is mainly used to appropriately extend the life cycle of fresh plants, and arrange the supply of fresh plants according to sales volume to ensure long-term uninterrupted supply. In production practice, there is no precedent for large-scale seedling cultivation using asexual reproduction techniques such as cuttings and tissue culture.

Due to the poor natural reproductive capacity of male stone pine, we use tissue culture technology to quickly propagate seedlings. Because tissue culture seedlings grow in a sterile state, they cannot make good use of their roots to absorb nutrients in the soil when transplanting and growing slowly. The male stone pine is cultivated by using the planted orchid base material (the base material is more suitable for the propagation of endophytic fungi) and the artificially cultivated endophytic fungus mixed substrate. After the fungus invades the roots of the male stone pine, a symbiotic mycorrhizal system can be formed, which is the growth of the male stone pine. The development provides relevant nutrients. The present invention uses the seedlings of wild male stone pine through tissue culture, and uses planted orchid base material to add artificially cultivated endophytic fungus mixed substrate to cultivate male stone pine, which can obviously promote plant growth and weight gain and improve product quality. , so as to better develop and utilize the medicinal value of male stone pine.

Contents of the invention

The object of the present invention is to provide a kind of method that utilizes orchid root fungi to cultivate male stone pine. The present invention uses wild male stone pine seedlings through tissue culture, adopts planted orchid base material and adds artificially cultivated endophytic fungus mixed matrix to cultivate male stone pine, which can It obviously promotes the increase of plant height and weight and improves the quality of products, so as to better develop and utilize the medicinal value of male stone pine.

To achieve the above object, the present invention adopts the following technical solutions:

Utilize the method for cultivating male stone pine with orchid rhizome, comprising the steps:

(1) Isolation and cultivation of symbiotic bacteria; the orchid mycorrhizae were cleaned with tap water, and the surface was sterilized with alcohol with a volume fraction of 75%. Extract the hyphae and move them into the same plate medium for purification and culture; culture the mycelia at 25-30°C for 7-8 days to cover the plate medium, cut the mycelium pieces on the plate and inoculate them into mixed bacteria culture bags; at 24-28°C Cultivate for 35-40 days until the hyphae grow through the fungus bag; mix the mycelia through the fungus bag with the base material that has been planted with orchids, and use it as the male stone pine cultivation substrate;

(2) Planting and cultivation of male stone pine; When the male stone pine tissue culture seedlings are 3-3.5cm long, take them out of the culture bottle, rinse the root culture medium with tap water; spread a layer of 1cm yellow soil on the male stone pine cultivation medium, wash the The male stone pine tissue culture seedlings were planted on the substrate, watered thoroughly and placed in a place without direct sunlight, and kept at 85% humidity; after 50-60 days of cultivation, nodular mycorrhizae were clearly seen on the roots.

The composition of the mixed bacteria culture bag is: 87-88% of sawdust, 2-3% of corn flour, 0.5-1% of lime powder, 7-8% of wheat bran, and 0.5-1% of sucrose. In the step (1), the weight ratio of the mycelia-grown fungus bag mixed with the orchid-planted base material is 1:30.

The composition of the fungal plate culture medium is: peptone 5.0g/L, beef extract 3.0g/L, agar 20.0g/L.

The advantages of the present invention are:

The present invention uses the seedlings of wild male stone pine through tissue culture, adopts planted orchid base material (currently large-scale planting of oncidium mainly uses a mixed matrix of small stones and bark (Figure 5), and the planting process needs to replace the base material regularly) to add artificial Cultivation of male stone pine with mixed substrate of cultured endophytic fungi. In this way, the present invention can make full use of the waste to plant the male stone pine, greatly reducing the planting cost. Effective rapid propagation and large-scale cultivation of male stone pine, so as to better develop and utilize the medicinal value of male stone pine.

The invention realizes rapid propagation and high-efficiency cultivation of seedlings under sterile conditions, with a survival rate of more than 90%, and the weight of a single plant after planting for 120 days is about 30% higher than that of ordinary substrates. It overcomes the phenomenon that common substrates are prone to rotten roots, thereby solving the problem of the lack of wild resources in nature caused by the poor natural reproductive capacity of male stone pine. To better develop and utilize this Chinese herbal medicine with a long history of application and definite efficacy fills up the technical research on asexual reproduction technology and high-efficiency cultivation. Open up a new model for the artificial cultivation of male stone pine.

Description of drawings

Figure 1 Endophytic fungal hyphae.

Figure 2. Endophytic fungi can be seen in the mycorrhizal sections of male Lycopodium pine.

Figure 3 Mycorrhizae of male Lycopodium.

Figure 4. Male stone pine cultivated with mixed substrates.

Figure 5 Oncidium planted in a mixed substrate of small stones and bark.

Figure 6 is covered with a layer of loess sublayer mixed matrix.

Detailed ways

Example 1

Utilize the method for cultivating male stone pine with orchid rhizome, comprising the steps:

(1) Isolation and cultivation of symbiotic bacteria; the orchid mycorrhizae were cleaned with tap water, and the surface was sterilized with alcohol with a volume fraction of 75%. Extract the mycelium and transfer it to the same plate medium for purification and culture; culture at 25°C for 7 days to cover the plate medium, cut the mycelium block on the plate and inoculate it into the mixed bacteria culture bag; cultivate at 24°C for 35 days until the bacteria Mycelium penetrating bacteria bag; mix mycelia penetrating bacteria bag with the base material planted with orchids (1:30), as the cultivation substrate of male stone pine;

(2) Planting and cultivation of male stone pine; When the male stone pine tissue culture seedlings are 3 cm long, take them out from the culture bottle, rinse the root culture medium with tap water; spread a layer of 1 cm yellow soil on the male stone pine cultivation substrate, and put the washed male stone pine The tissue culture seedlings were planted on the matrix, watered thoroughly and placed in a place without direct sunlight, and kept at 85% humidity; after 50 days of cultivation, nodular mycorrhizae were clearly seen on the roots.

The composition of the mixed bacteria culture bag is: 87% of sawdust, 3% of corn flour, 1% of lime powder, 8% of wheat bran, and 1% of sucrose. In the step (1), the weight ratio of the mycelia-grown fungus bag mixed with the orchid-planted base material is 1:30.

The composition of the fungal plate culture medium is: peptone 5.0g/L, beef extract 3.0g/L, agar 20.0g/L.

The invention realizes rapid propagation and high-efficiency cultivation of seedlings under sterile conditions, with a survival rate of 96%, and the weight of a single plant after planting for 120 days is 35% higher than that of ordinary substrates.

Example 2

Utilize the method for cultivating male stone pine with orchid rhizome, comprising the steps:

(1) Isolation and cultivation of symbiotic bacteria; the orchid mycorrhizae were cleaned with tap water, and the surface was sterilized with alcohol with a volume fraction of 75%. Extract the hyphae and transfer them to the same plate medium for purification and culture; culture at 30°C for 8 days to cover the plate medium, cut the mycelium pieces on the plate and inoculate them into mixed bacteria culture bags; cultivate at 28°C for 40 days until the bacteria Mycelium penetrating bacteria bag; mix mycelia penetrating bacteria bag with the base material planted with orchids (1:30), as the cultivation substrate of male stone pine;

(2) Planting and cultivation of male stone pine; when the male stone pine tissue culture seedlings are 3.5 cm long, take them out from the culture bottle, rinse the root culture medium with tap water; spread a layer of 1 cm yellow soil on the male stone pine cultivation medium, and put the washed male stone pine Lycopodium tissue culture seedlings were planted on the substrate, watered thoroughly and placed in a place without direct sunlight, and kept at 85% humidity; after 60 days of cultivation, nodular mycorrhizae were clearly seen at the roots.

The composition of the mixed bacteria culture bag is: 88% of sawdust, 2% of corn flour, 0.5% of lime powder, 7% of wheat bran, and 0.5% of sucrose. In the step (1), the weight ratio of the mycelia-grown fungus bag mixed with the orchid-planted base material is 1:30.

The composition of the fungal plate culture medium is: peptone 5.0g/L, beef extract 3.0g/L, agar 20.0g/L.

The invention realizes rapid propagation and high-efficiency cultivation of seedlings under sterile conditions, with a survival rate of 96%, and the weight of a single plant after planting for 120 days is 35% higher than that of ordinary substrates.

Example 3

Utilize the method for cultivating male stone pine with orchid rhizome, comprising the steps:

(1) Isolation and cultivation of symbiotic bacteria; the orchid mycorrhizae were cleaned with tap water, and the surface was sterilized with alcohol with a volume fraction of 75%. Extract the hyphae and transfer them to the same plate medium for purification and culture; culture at 28°C for 8 days to cover the plate medium, cut the mycelium pieces on the plate and inoculate them into the mixed bacteria culture bag; cultivate at 25°C for 38 days until the bacteria Mycelium penetrating bacteria bag; mix mycelia penetrating bacteria bag with the base material planted with orchids (1:30), as the cultivation substrate of male stone pine;

(2) Planting and cultivation of male stone pine; when the male stone pine tissue culture seedlings are 3.5 cm long, take them out from the culture bottle, rinse the root culture medium with tap water; spread a layer of 1 cm yellow soil on the male stone pine cultivation medium, and put the washed male stone pine Lycopodium tissue culture seedlings are planted on the substrate, watered thoroughly and placed in a place without direct sunlight, and the humidity is maintained at 85%. After 55 days of cultivation, nodular mycorrhizae can be clearly seen at the roots.

The composition of the mixed bacteria culture bag is: 87% of sawdust, 3% of corn flour, 1% of lime powder, 8% of wheat bran, and 1% of sucrose. In the step (1), the weight ratio of the mycelia-grown fungus bag mixed with the orchid-planted base material is 1:30.

The composition of the fungal plate culture medium is: peptone 5.0g/L, beef extract 3.0g/L, agar 20.0g/L.

The invention realizes rapid propagation and high-efficiency cultivation of seedlings under sterile conditions, with a survival rate of 96%, and the weight of a single plant after planting for 120 days is 35% higher than that of ordinary substrates.

The above descriptions are only preferred embodiments of the present invention, and all equivalent changes and modifications made according to the scope of the patent application of the present invention shall fall within the scope of the present invention.

Claims (4)

1. utilize the method for orchid root fungus cultivation male stone pine, it is characterized in that: described method comprises the steps: (1) Isolation and cultivation of symbiotic bacteria; the orchid mycorrhizae were cleaned with tap water, and the surface was sterilized with alcohol with a volume fraction of 75%. Extract the hyphae and move them into the same plate medium for purification and culture; culture the mycelia at 25-30°C for 7-8 days to cover the plate medium, cut the mycelium pieces on the plate and inoculate them into mixed bacteria culture bags; at 24-28°C Cultivate for 35-40 days until the hyphae grow through the bacteria bag; mix the mycelium through the bacteria bag with the base material that has been planted with orchids, and use it as the male stone pine cultivation substrate; (2) Planting and cultivation of male stone pine; When the male stone pine tissue culture seedling grows to 3-3.5cm, take it out from the culture bottle, rinse the root medium with tap water; spread a layer of 1cm yellow soil on the male stone pine cultivation medium, and wash the The male stone pine tissue culture seedlings were planted on the substrate, watered thoroughly and placed in a place without direct sunlight, and kept at 85% humidity; after 50-60 days of cultivation, nodular mycorrhizae were clearly seen on the roots. 2. The method of utilizing orchid root fungus to cultivate male stone pine according to claim 1, characterized in that: the composition of the mixed bacteria culture bag is: sawdust 87-88%, corn flour 2-3%, lime powder 0.5% -1%, wheat bran 7-8%, sucrose 0.5-1%. 3. The method for cultivating male stone pine with orchid root fungus according to claim 1, characterized in that: in step (1), the weight ratio of the mycelia-grown fungus bag mixed with the orchid-planted base material is 1:30 . 4. the method for utilizing orchid root fungus to cultivate male stone pine according to claim 1, is characterized in that: the composition of described fungus plate culture medium is: peptone 5.0g/L, beef extract 3.0g/L, agar 20.0g/L L.