Method for artificially breeding cibotium barometz by using spores

Abstract

The invention belongs to the technical field of cibotium barometz breeding, and particularly relates to a method for artificially breeding cibotium barometz by using spores, which comprises the following components in parts by weight: 100 parts of field soil, 20-25 parts of sawdust, 10-15 parts of perlite, 10-12 parts of plant ash, 0.05-0.15 part of manganese sulfate and 0.3-0.5 part of sodium selenite; spores germinate, and the environment temperature is 24-26 DEG C; and transplanting the germinated seedlings into a greenhouse for cultivation. According to the method, high spore germination rate and seedling survival rate are ensured, and the transplanted seedlings can grow smoothly.

Description

Technical Field

[0001] The invention belongs to the technical field of golden retriever breeding, and particularly relates to a method for artificially breeding golden retrievers by utilizing spores. Background Art

[0002] Cibotium barometz (L.) J.Sm. is a tree fern of the Cibotidae family and Cibotium genus. It occupies an important position in the ecosystem. At present, the existing technology for the study of Cibotium barometz presents a multi-dimensional trend, mainly including: (1) morphological diversity research, the purpose of which is to reveal the law of morphological variation of Cibotium barometz in different geographical regions and ecological environments, and provide a solid morphological basis for the classification, identification and evolutionary research of species; (2) community classification research, focusing on the distribution of Cibotium barometz in different ecological communities and its relationship with surrounding plants, aiming to clarify the suitable ecological niche of Cibotium barometz and provide scientific guidance for the protection and restoration of its habitat; (3) genetic diversity research, focusing on the distribution of Cibotium barometz in different ecological communities and its relationship with surrounding plants, and providing scientific guidance for the protection and restoration of its habitat; (4) genetic diversity research, focusing on the distribution of Cibotium barometz in different ecological communities and its relationship with surrounding plants, and providing scientific guidance for the protection and restoration of its habitat; (5) genetic diversity research, focusing on the distribution of Cibotium barometz in different ecological communities and its relationship with surrounding plants, and (6) genetic diversity research, focusing on the distribution of Cibotium barometz in different ecological communities and its relationship with surrounding plants, and (7) genetic diversity research, focusing on the distribution of Cibotium barometz in different ecological communities and its relationship with surrounding plants, and (8) genetic diversity research, focusing on the distribution of Cibotium barometz in different ecological communities and its relationship with surrounding plants, and (9) genetic diversity research, focusing on the distribution of Cibotium barometz in different ecological communities and its relationship with surrounding plants, and (10) genetic diversity research, focusing on the distribution of Cibotium barometz in different ecological communities and (11) genetic diversity research, focusing on the distribution of Cibotium barometz in different ecological communities and its relationship with surrounding plants, and (12) genetic diversity research, focusing on the distribution of Cibotium barometz in different ecological communities and (13) genetic diversity research, focusing (4) Allelopathic research, focusing on the effects of chemical substances produced by the plants themselves on the growth and development of other organisms around them; (5) Breeding research, such as the study of the stages of gametophyte development, tracking the stages of gametophyte development from spore germination to gametophyte maturation and fertilization, studying the needs and responses of gametophytes to environmental factors at different developmental stages, and providing a theoretical basis for optimizing gametophyte development conditions during artificial breeding. All of the above studies require the use of plants, such as roots, stems, and leaves, as research materials, so the demand for a large number of plants is high.

[0003] However, the Golden Retriever's reproduction process is significantly affected by season and temperature. Seasonal changes and temperature fluctuations make it difficult to predict the timing of Golden Retriever reproduction, resulting in a chronically low reproduction coefficient. This situation makes it difficult to obtain large quantities of seedlings in natural conditions, severely limiting the expansion of the Golden Retriever population and its distribution range.

[0004] Although there are studies in the prior art on obtaining golden dog seedlings using spore propagation, golden dog spore propagation is extremely sensitive to environmental conditions. Factors such as soil pH, fertility, and temperature can adversely affect the implantation and germination of spores, resulting in a low spore germination rate and restricting the breeding of golden dog. Summary of the Invention

[0005] In order to solve the above technical problems, the present invention provides a method for artificially breeding golden retrievers using spores.

[0006] The object of the present invention is to provide a method for artificially breeding golden retriever dogs using spores, comprising:

[0007] Substrate preparation steps: The substrate formula, by weight, is as follows: 100 parts garden soil, 20-25 parts sawdust, 10-15 parts perlite, 10-12 parts wood ash, 0.05-0.15 parts manganese sulfate, and 0.3-0.5 parts sodium selenite. Garden soil, sawdust, and wood ash have good air permeability, drainage, and moderate water retention, making them ideal substrates for dogwood spore germination. Garden soil, sawdust, and wood ash also provide a carbon source and other nutrients. Perlite is breathable. Sodium selenite provides selenium nutrition and, in combination with manganese sulfate, stimulates dogwood spore germination.

[0008] The spores germinate, and the environmental temperature for spore germination is 24°C to 26°C; this temperature range is the optimal temperature for spore germination of golden dog, which can ensure that various physiological and biochemical reactions in the spores are carried out efficiently.

[0009] Seedling cultivation: Germinated seedlings are transplanted into greenhouses for cultivation. The greenhouses provide a relatively stable temperature, humidity, and light environment for the seedlings, reducing the impact of environmental fluctuations on their growth.

[0010] Preferably, in the above-mentioned method of artificially breeding golden retrievers using spores, the matrix formula is as follows: 100 parts of garden soil, 20 parts of sawdust, 10 parts of perlite, 10 parts of plant ash, 0.1 parts of manganese sulfate, and 0.4 parts of sodium selenite.

[0011] Preferably, in the above method of artificially breeding golden retrievers using spores, the sawdust is poplar sawdust powder.

[0012] Preferably, in the above-mentioned method for artificially breeding golden retrievers using spores, spore germination is performed indoors under a lighting condition of 2500-3000 lux for 10-12 hours. This avoids burns to the spores from excessive light while meeting their photosynthesis requirements, simulating the light rhythm in the natural environment and promoting normal spore germination and early growth.

[0013] Preferably, in the above method of artificially breeding golden retrievers using spores, the indoor environmental humidity is controlled at 70% to 80%.

[0014] Preferably, in the above-mentioned method of artificially breeding golden retrievers using spores, the temperature of the greenhouse is 24° C. to 26° C., and the humidity is 70% to 75%.

[0015] Preferably, in the above method of artificially breeding golden retriever using spores, the soil water holding capacity is maintained at 60% to 65%.

[0016] The present invention provides an application of a matrix in improving the artificial breeding efficiency of golden retrievers. The matrix is ​​the above-mentioned matrix.

[0017] Preferably, improving the artificial breeding efficiency of golden dog refers to improving the spore germination rate, the seedling acquisition rate, the survival rate of the seedlings after transplanting, and promoting the development of the seedlings.

[0018] Preferably, the promoting seedling development refers to accelerating the production of new leaves and increasing leaf length and leaf width.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] The present invention greatly improves the spore germination rate through precise control of the matrix formula (100 parts of garden soil, 20 to 25 parts of sawdust, 10 to 15 parts of perlite, 10 to 12 parts of wood ash, 0.05 to 0.15 parts of manganese sulfate, and 0.3 to 0.5 parts of sodium selenite), the germination environment, and the seedling cultivation process. Manganese sulfate and sodium selenite are added to the matrix formula, and during the spore germination stage, the environmental temperature is controlled at 24°C to 26°C, so that the spores can break through dormancy in a shorter time and start the germination process. During the seedling cultivation stage, cultivation is carried out in a greenhouse, which ensures a high spore germination rate and seedling survival rate, and the seedlings after transplanting can grow smoothly, effectively improving the survival rate of later forest planting (up to 95%).

[0021] The present invention can produce a large number of uniformly grown seedlings of uniform physiological age, ensuring consistency in the initial stages of seedling development. During subsequent seedling cultivation and transplanting, uniform environmental settings and management measures further promote uniform seedling growth. This characteristic makes the technology well-suited for large-scale seedling cultivation, meeting market demand for large quantities of high-quality golden retriever seedlings and promoting the development of the artificial breeding industry for golden retrievers. DETAILED DESCRIPTION

[0022] In order to enable those skilled in the art to better understand and implement the technical solution of the present invention, the present invention is further described below with reference to specific embodiments.

[0023] In the description of the present invention, unless otherwise specified, all reagents used are commercially available and all methods used are conventional techniques in the art.

[0024] The sources of raw materials used in the following examples and controls are as follows:

[0025] (1) The spores of Golden Dog were collected and preserved according to the following method: The collection site was Lushan City, Jiujiang City, Jiangxi Province. Mature and healthy Golden Dog plants were selected. The sign of maturity was that the spore sacs were dark brown and plump. At this time, the spores were fully developed and had good germination ability. Use sterilized scissors to collect small leaves with mature spores and place them in a clean paper bag. The paper bag has good air permeability, which can prevent the spores from being lost and ensure air circulation to prevent the spores from deteriorating. The paper bag containing the spore sample was brought back to the laboratory and left to dry in the shade under the laboratory environment. After the spores naturally fell from the leaves, they were collected and then stored at 4℃ for later use.

[0026] (2) Garden soil, collected from Guling Town, Lushan City, Jiujiang City, Jiangxi Province.

[0027] (3) Sawdust, purchased from Caoxian Shenhang Wood Industry Co., Ltd., is premium pure poplar sawdust powder.

[0028] (4) Perlite, purchased from Lingshou County Chenyang Mineral Products Co., Ltd., with a particle size of 1 mm to 3 mm.

[0029] (5) Wood ash, purchased from Hebei Xiong'an Xinying Technology Co., Ltd., pH 8.5-9, water absorption 40%, dark brown.

[0030] In order to explore the effects of different amounts of manganese sulfate added on the artificial breeding efficiency of golden retrievers, we set up Examples 1 to 3, a blank control, and a single-factor control 1.

[0031] Example 1

[0032] A method for artificially breeding golden retrievers using spores, comprising:

[0033] Step 1, Matrix Preparation

[0034] The matrix formula is as follows, in parts by weight: 100 parts of garden soil, 20 parts of sawdust, 10 parts of perlite, 10 parts of plant ash, 0.05 parts of manganese sulfate, and 0.3 parts of sodium selenite. The raw materials are thoroughly mixed to obtain the matrix.

[0035] Before use, sterilize the substrate, for example, at 121°C for 20 minutes under high-temperature and high-pressure conditions to completely kill pathogens, insect eggs, and other harmful microorganisms in the soil, creating a sterile environment ideal for spore germination. After sterilization, place the substrate in a nutrient cup, water thoroughly, and set aside.

[0036] Step 2, spore germination

[0037] Golden dog spores were rinsed three times with sterile water to remove dust and pathogens. After rinsing, the spores were evenly spread on the surface of the substrate in nutrient cups. Each nutrient cup was inoculated with 50 spores. The nutrient cups were 22.3 cm long, 15.3 cm wide, and 7.9 cm high, for a total of 300 spores. The spores were covered with a transparent plastic lid to maintain moisture and incubated indoors. Each treatment was replicated three times. During spore germination, the ambient temperature was set at 25°C, and the lighting conditions were set at 2500 lux for 10 hours of light and 14 hours of darkness. The indoor humidity was controlled at 70%.

[0038] On the 30th day, the number of germinated spores was counted by microscope observation. The sign of spore germination was that the prothallus broke through the spore wall and extended green filaments and lamellae. The seedling yield rate was calculated as follows:

[0039] Spore germination rate = (number of spores that have germinated into sporophytes / 300) × 100%.

[0040] On the 90th day, observe and record the number of seedlings at the two-leaf, one-heart stage, and calculate the seedling yield rate:

[0041] Seedling acquisition rate = (number of seedlings at the two-leaf, one-heart stage / 300) × 100%.

[0042] Step 3: Seedling cultivation

[0043] After a period of cultivation, when the seedlings have grown to 3 leaves and a height of 2 cm, they are capable of independent life and can be transplanted to a greenhouse. After transplanting, water the soil regularly every week to maintain a water holding capacity of 60%, but avoid stagnant water to prevent root rot.

[0044] The formula of the transplanting substrate was the same as that in step 1. Each treatment group was repeated for 100 plants. The growth status of the seedlings was recorded within one month, and the average value of each index was taken.

[0045] The temperature of the greenhouse was controlled at 25°C and the humidity was 70%.

[0046] The light transmittance of the greenhouse is controlled at 50%, and the greenhouse provides a diffuse light environment.

[0047] Example 2

[0048] A method for artificially breeding golden retrievers using spores is basically the same as the operation of Example 1, except that the matrix formula is as follows: 100 parts of garden soil, 20 parts of sawdust, 10 parts of perlite, 10 parts of wood ash, 0.1 part of manganese sulfate, and 0.3 part of sodium selenite.

[0049] Specifically, the method of Example 2 includes:

[0050] Step 1, Matrix Preparation

[0051] The matrix formula is as follows, in parts by weight: 100 parts of garden soil, 20 parts of sawdust, 10 parts of perlite, 10 parts of plant ash, 0.1 parts of manganese sulfate, and 0.3 parts of sodium selenite. The raw materials are thoroughly mixed to obtain the matrix.

[0052] Before use, sterilize the substrate, for example, at 121°C for 20 minutes under high-temperature and high-pressure conditions to completely kill pathogens, insect eggs, and other harmful microorganisms in the soil, creating a sterile environment ideal for spore germination. After sterilization, place the substrate in a nutrient cup, water thoroughly, and set aside.

[0053] Step 2, spore germination

[0054] Golden dog spores were rinsed three times with sterile water to remove dust and pathogens. After rinsing, the spores were evenly spread on the surface of the substrate in nutrient cups. Each nutrient cup was inoculated with 50 spores. The nutrient cups were 22.3 cm long, 15.3 cm wide, and 7.9 cm high, for a total of 300 spores. The spores were covered with a transparent plastic lid to maintain moisture and incubated indoors. Each treatment was replicated three times. During spore germination, the ambient temperature was set at 25°C, and the lighting conditions were set at 2500 lux for 10 hours of light and 14 hours of darkness. The indoor humidity was controlled at 70%.

[0055] On the 30th day, the number of germinated spores was observed and counted under a microscope. On the 90th day, the number of seedlings at the two-leaf, one-heart stage was observed and recorded. The spore germination rate and seedling acquisition rate were calculated according to the formula in Example 1.

[0056] Step 3: Seedling cultivation

[0057] After a period of cultivation, when the seedlings have grown to three leaves and a height of 2 cm, they are capable of independent life and are ready for transplanting. At this point, they are transplanted into a greenhouse. After transplanting, water the soil regularly weekly to maintain a water holding capacity of 60%, but avoid stagnant water to prevent root rot.

[0058] The formula of the transplanting substrate was the same as that in step 1. Each treatment group was repeated for 100 plants. The growth status of the seedlings was recorded within one month, and the average value of each index was taken.

[0059] The temperature of the greenhouse was controlled at 25°C and the humidity was 70%.

[0060] The light transmittance of the greenhouse is controlled at 50%, and the greenhouse provides a diffuse light environment.

[0061] Example 3

[0062] A method for artificially breeding golden retrievers using spores is basically the same as the operation of Example 1, except that the matrix formula is as follows: 100 parts of garden soil, 20 parts of sawdust, 10 parts of perlite, 10 parts of plant ash, 0.15 parts of manganese sulfate, and 0.3 parts of sodium selenite.

[0063] Specifically, the method of Example 3 includes:

[0064] Step 1, Matrix Preparation

[0065] The matrix formula is as follows, in parts by weight: 100 parts of garden soil, 20 parts of sawdust, 10 parts of perlite, 10 parts of plant ash, 0.15 parts of manganese sulfate, and 0.3 parts of sodium selenite. The raw materials are thoroughly mixed to obtain the matrix.

[0066] Before use, sterilize the substrate, for example, at 121°C for 20 minutes under high-temperature and high-pressure conditions to completely kill pathogens, insect eggs, and other harmful microorganisms in the soil, creating a sterile environment ideal for spore germination. After sterilization, place the substrate in a nutrient cup, water thoroughly, and set aside.

[0067] Step 2, spore germination

[0068] Golden dog spores were rinsed three times with sterile water to remove dust and pathogens. After rinsing, the spores were evenly spread on the surface of the substrate in nutrient cups. Each nutrient cup was inoculated with 50 spores. The nutrient cups were 22.3 cm long, 15.3 cm wide, and 7.9 cm high, for a total of 300 spores. The spores were incubated indoors, with each treatment replicated three times. During spore germination, the ambient temperature was set at 25°C, and the lighting conditions were set at 2500 lux for 10 hours of light and 14 hours of darkness. The indoor humidity was controlled at 70%.

[0069] On the 30th day, the number of germinated spores was observed and counted under a microscope. On the 90th day, the number of seedlings at the two-leaf, one-heart stage was observed and recorded. The spore germination rate and seedling acquisition rate were calculated according to the formula in Example 1.

[0070] Step 3: Seedling cultivation

[0071] After a period of growth, when the seedlings have grown to three leaves and a height of 2 cm, they are capable of independent life and are ready for transplanting to a greenhouse. After transplanting, the seedlings should be placed in the greenhouse for cultivation. Water regularly weekly to maintain the soil's water holding capacity at 60%, but avoid stagnant water to prevent root rot.

[0072] The formula of the transplanting substrate was the same as that in step 1. Each treatment group was repeated for 100 plants. The growth status of the seedlings was recorded within one month, and the average value of each index was taken.

[0073] The temperature of the greenhouse was controlled at 25°C and the humidity was 70%.

[0074] The light transmittance of the greenhouse is controlled at 50%, and the greenhouse provides a diffuse light environment.

[0075] Blank control

[0076] A method for artificially breeding golden retrievers using spores is basically the same as the operation of Example 1, except that the matrix formula is as follows: 100 parts of garden soil, 20 parts of sawdust, 10 parts of perlite, and 10 parts of wood ash.

[0077] Specifically, the blank control method includes:

[0078] Step 1, Matrix Preparation

[0079] The matrix formula is as follows, in parts by weight: 100 parts of garden soil, 20 parts of sawdust, 10 parts of perlite, and 10 parts of plant ash. The raw materials are thoroughly mixed to obtain the matrix.

[0080] Before use, sterilize the substrate, for example, at 121°C for 20 minutes under high-temperature and high-pressure conditions to completely kill pathogens, insect eggs, and other harmful microorganisms in the soil, creating a sterile environment ideal for spore germination. After sterilization, place the substrate in a nutrient cup, water thoroughly, and set aside.

[0081] Step 2, spore germination

[0082] Golden dog spores were rinsed three times with sterile water to remove dust and pathogens. After rinsing, the spores were evenly spread on the surface of the substrate in nutrient cups. Each nutrient cup was inoculated with 50 spores. The nutrient cups were 22.3 cm long, 15.3 cm wide, and 7.9 cm high, for a total of 300 spores. The spores were covered with a transparent plastic lid to maintain moisture and incubated indoors. Each treatment was replicated three times. During spore germination, the ambient temperature was set at 25°C, and the lighting conditions were set at 2500 lux for 10 hours of light and 14 hours of darkness. The indoor humidity was controlled at 70%.

[0083] On the 30th day, the number of germinated spores was observed and counted under a microscope. On the 90th day, the number of seedlings at the two-leaf, one-heart stage was observed and recorded. The spore germination rate and seedling acquisition rate were calculated according to the formula in Example 1.

[0084] Step 3: Seedling cultivation

[0085] After a period of growth, when the seedlings have grown to three leaves and a height of 2 cm, they are capable of independent life and can be transplanted. After transplanting, the seedlings should be placed in a greenhouse for cultivation. Water regularly weekly to maintain the soil's water holding capacity at 60%, but avoid stagnant water to prevent root rot.

[0086] The formula of the transplanting substrate was the same as that in step 1. Each treatment group was repeated for 100 plants. The growth status of the seedlings was recorded within one month, and the average value of each index was taken.

[0087] The temperature of the greenhouse was controlled at 25°C and the humidity was 70%.

[0088] The light transmittance of the greenhouse is controlled at 50%, and the greenhouse provides a diffuse light environment.

[0089] Single factor control 1

[0090] A method for artificially breeding golden retrievers using spores is basically the same as the operation of Example 1, except that the matrix formula is as follows: 100 parts of garden soil, 20 parts of sawdust, 10 parts of perlite, 10 parts of plant ash, and 0.05 parts of manganese sulfate.

[0091] Specifically, the method of single factor control 1 includes:

[0092] Step 1, Matrix Preparation

[0093] The matrix formula is as follows, in parts by weight: 100 parts of garden soil, 20 parts of sawdust, 10 parts of perlite, 10 parts of plant ash, and 0.05 parts of manganese sulfate. The raw materials are thoroughly mixed to obtain the matrix.

[0094] Before use, sterilize the substrate, for example, at 121°C for 20 minutes under high-temperature and high-pressure conditions to completely kill pathogens, insect eggs, and other harmful microorganisms in the soil, creating a sterile environment ideal for spore germination. After sterilization, place the substrate in a nutrient cup, water thoroughly, and set aside.

[0095] Step 2, spore germination

[0096] Golden dog spores were rinsed three times with sterile water to remove dust and pathogens. After rinsing, the spores were evenly spread on the surface of the substrate in nutrient cups. Each nutrient cup was inoculated with 50 spores. The nutrient cups were 22.3 cm long, 15.3 cm wide, and 7.9 cm high, for a total of 300 spores. The spores were covered with a transparent plastic lid to maintain moisture and incubated indoors. Each treatment was replicated three times. During spore germination, the ambient temperature was set at 25°C, and the lighting conditions were set at 2500 lux for 10 hours of light and 14 hours of darkness. The indoor humidity was controlled at 70%.

[0097] On the 30th day, the number of germinated spores was observed and counted under a microscope. On the 90th day, the number of seedlings at the two-leaf, one-heart stage was observed and recorded. The spore germination rate and seedling acquisition rate were calculated according to the formula in Example 1.

[0098] Step 3: Seedling cultivation

[0099] After a period of growth, when the seedlings have grown to three leaves and a height of 2 cm, they are capable of independent life and can be transplanted. After transplanting, the seedlings should be placed in a greenhouse for cultivation. Water regularly weekly to maintain the soil's water holding capacity at 60%, but avoid stagnant water to prevent root rot.

[0100] The formula of the transplanting substrate was the same as that in step 1. Each treatment group was repeated for 100 plants. The growth status of the seedlings was recorded within one month, and the average value of each index was taken.

[0101] The temperature of the greenhouse was controlled at 25°C and the humidity was 70%.

[0102] The light transmittance of the greenhouse is controlled at 50%, and the greenhouse provides a diffuse light environment.

[0103] In order to explore the effects of different sodium selenite dosages on the artificial breeding efficiency of golden retrievers, we set up Example 2, Example 4-Example 5, blank control and single factor control 2. Among them, Example 4-Example 5 and single factor control 2 are as follows.

[0104] Example 4

[0105] A method for artificially breeding golden retrievers using spores is basically the same as the operation of Example 1, except that the matrix formula is as follows: 100 parts of garden soil, 20 parts of sawdust, 10 parts of perlite, 10 parts of wood ash, 0.1 part of manganese sulfate, and 0.4 part of sodium selenite.

[0106] Specifically, the method of Example 4 includes:

[0107] Step 1, Matrix Preparation

[0108] The matrix formula is as follows, in parts by weight: 100 parts of garden soil, 20 parts of sawdust, 10 parts of perlite, 10 parts of plant ash, 0.1 parts of manganese sulfate, and 0.4 parts of sodium selenite. The raw materials are thoroughly mixed to obtain the matrix.

[0109] Before use, sterilize the substrate, for example, at 121°C for 20 minutes under high-temperature and high-pressure conditions to completely kill pathogens, insect eggs, and other harmful microorganisms in the soil, creating a sterile environment ideal for spore germination. After sterilization, place the substrate in a nutrient cup, water thoroughly, and set aside.

[0110] Step 2, spore germination

[0111] Golden dog spores were rinsed three times with sterile water to remove dust and pathogens. After rinsing, the spores were evenly spread on the surface of the substrate in nutrient cups. Each nutrient cup was inoculated with 50 spores. The nutrient cups were 22.3 cm long, 15.3 cm wide, and 7.9 cm high, for a total of 300 spores. The spores were covered with a transparent plastic lid to maintain moisture and incubated indoors. Each treatment was replicated three times. During spore germination, the ambient temperature was set at 25°C, and the lighting conditions were set at 2500 lux for 10 hours of light and 14 hours of darkness. The indoor humidity was controlled at 70%.

[0112] On the 30th day, the number of germinated spores was observed and counted under a microscope. On the 90th day, the number of seedlings at the two-leaf, one-heart stage was observed and recorded. The spore germination rate and seedling acquisition rate were calculated according to the formula in Example 1.

[0113] Step 3: Seedling cultivation

[0114] After a period of growth, when the seedlings have grown to three leaves and a height of 2 cm, they are capable of independent life and can be transplanted. After transplanting, the seedlings should be placed in a greenhouse for cultivation. Water regularly weekly to maintain the soil's water holding capacity at 60%, but avoid stagnant water to prevent root rot.

[0115] The formula of the transplanting substrate was the same as that in step 1. Each treatment group was repeated for 100 plants. The growth status of the seedlings was recorded within one month, and the average value of each index was taken.

[0116] The temperature of the greenhouse was controlled at 25°C and the humidity was 70%.

[0117] The light transmittance of the greenhouse is controlled at 50%, and the greenhouse provides a diffuse light environment.

[0118] Example 5

[0119] A method for artificially breeding golden retrievers using spores is basically the same as the operation of Example 1, except that the matrix formula is as follows: 100 parts of garden soil, 20 parts of sawdust, 10 parts of perlite, 10 parts of plant ash, 0.1 part of manganese sulfate, and 0.5 part of sodium selenite.

[0120] Specifically, the method of Example 5 includes:

[0121] Step 1, Matrix Preparation

[0122] The matrix formula is as follows, in parts by weight: 100 parts of garden soil, 20 parts of sawdust, 10 parts of perlite, 10 parts of plant ash, 0.1 parts of manganese sulfate, and 0.5 parts of sodium selenite. The raw materials are thoroughly mixed to obtain the matrix.

[0123] Before use, sterilize the substrate, for example, at 121°C for 20 minutes under high-temperature and high-pressure conditions to completely kill pathogens, insect eggs, and other harmful microorganisms in the soil, creating a sterile environment ideal for spore germination. After sterilization, place the substrate in a nutrient cup, water thoroughly, and set aside.

[0124] Step 2, spore germination

[0125] Golden dog spores were rinsed three times with sterile water to remove dust and pathogens. After rinsing, the spores were evenly spread on the surface of the substrate in nutrient cups. Each nutrient cup was inoculated with 50 spores. The nutrient cups were 22.3 cm long, 15.3 cm wide, and 7.9 cm high, for a total of 300 spores. The spores were covered with a transparent plastic lid to maintain moisture and incubated indoors. Each treatment was replicated three times. During spore germination, the ambient temperature was set at 25°C, and the lighting conditions were set at 2500 lux for 10 hours of light and 14 hours of darkness. The indoor humidity was controlled at 70%.

[0126] On the 30th day, the number of germinated spores was observed and counted under a microscope. On the 90th day, the number of seedlings at the two-leaf, one-heart stage was observed and recorded. The spore germination rate and seedling acquisition rate were calculated according to the formula in Example 1.

[0127] Step 3: Seedling cultivation

[0128] After a period of growth, when the seedlings have grown to three leaves and a height of 2 cm, they are capable of independent life and can be transplanted. After transplanting, the seedlings should be placed in a greenhouse for cultivation. Water regularly weekly to maintain the soil's water holding capacity at 60%, but avoid stagnant water to prevent root rot.

[0129] The formula of the transplanting substrate was the same as that in step 1. Each treatment group was repeated for 100 plants. The growth status of the seedlings was recorded within one month, and the average value of each index was taken.

[0130] The temperature of the greenhouse was controlled at 25°C and the humidity was 70%.

[0131] The light transmittance of the greenhouse is controlled at 50%, and the greenhouse provides a diffuse light environment.

[0132] Single factor control 2

[0133] A method for artificially breeding golden retrievers using spores is basically the same as the operation of Example 1, except that the matrix formula is as follows: 100 parts of garden soil, 20 parts of sawdust, 10 parts of perlite, 10 parts of wood ash, and 0.3 parts of sodium selenite.

[0134] Specifically, the method of single factor control 2 includes:

[0135] Step 1, Matrix Preparation

[0136] The matrix formula is as follows, in parts by weight: 100 parts of garden soil, 20 parts of sawdust, 10 parts of perlite, 10 parts of plant ash, and 0.3 parts of sodium selenite. The raw materials are thoroughly mixed to obtain the matrix.

[0137] Before use, sterilize the substrate, for example, at 121°C for 20 minutes under high-temperature and high-pressure conditions to completely kill pathogens, insect eggs, and other harmful microorganisms in the soil, creating a sterile environment ideal for spore germination. After sterilization, place the substrate in a nutrient cup, water thoroughly, and set aside.

[0138] Step 2, spore germination

[0139] Golden dog spores were rinsed three times with sterile water to remove dust and pathogens. After rinsing, the spores were evenly spread on the surface of the substrate in nutrient cups. Each nutrient cup was inoculated with 50 spores. The nutrient cups were 22.3 cm long, 15.3 cm wide, and 7.9 cm high, for a total of 300 spores. The spores were covered with a transparent plastic lid to maintain moisture and incubated indoors. Each treatment was replicated three times. During spore germination, the ambient temperature was set at 25°C, and the lighting conditions were set at 2500 lux for 10 hours of light and 14 hours of darkness. The indoor humidity was controlled at 70%.

[0140] On the 30th day, the number of germinated spores was observed and counted under a microscope. On the 90th day, the number of seedlings at the two-leaf, one-heart stage was observed and recorded. The spore germination rate and seedling acquisition rate were calculated according to the formula in Example 1.

[0141] Step 3: Seedling cultivation

[0142] After a period of growth, when the seedlings have grown to three leaves and a height of 2 cm, they are capable of independent life and can be transplanted. After transplanting, the seedlings should be placed in a greenhouse for cultivation. Water regularly weekly to maintain the soil's water holding capacity at 60%, but avoid stagnant water to prevent root rot.

[0143] The formula of the transplanting substrate was the same as that in step 1. Each treatment group was repeated for 100 plants. The growth status of the seedlings was recorded within one month, and the average value of each index was taken.

[0144] The temperature of the greenhouse was controlled at 25°C and the humidity was 70%.

[0145] The light transmittance of the greenhouse is controlled at 50%, and the greenhouse provides a diffuse light environment.

[0146] In order to explore the effects of other matrix formula raw materials on the artificial breeding efficiency of golden retriever dogs, we set up Examples 4 and 6.

[0147] Example 6

[0148] A method for artificially breeding golden retrievers using spores is basically the same as the operation of Example 1, except that the matrix formula is as follows: 100 parts of garden soil, 25 parts of sawdust, 15 parts of perlite, 12 parts of plant ash, 0.1 part of manganese sulfate, and 0.4 part of sodium selenite.

[0149] Specifically, the method of Example 6 includes:

[0150] Step 1, Matrix Preparation

[0151] The matrix formula is as follows, in parts by weight: 100 parts of garden soil, 25 parts of sawdust, 15 parts of perlite, 12 parts of plant ash, 0.1 parts of manganese sulfate, and 0.4 parts of sodium selenite. The raw materials are thoroughly mixed to obtain the matrix.

[0152] Before use, sterilize the substrate, for example, at 121°C for 20 minutes under high-temperature and high-pressure conditions to completely kill pathogens, insect eggs, and other harmful microorganisms in the soil, creating a sterile environment ideal for spore germination. After sterilization, place the substrate in a nutrient cup, water thoroughly, and set aside.

[0153] Step 2, spore germination

[0154] Golden dog spores were rinsed three times with sterile water to remove dust and pathogens. After rinsing, the spores were evenly spread on the surface of the substrate in nutrient cups. Each nutrient cup was inoculated with 50 spores. The nutrient cups were 22.3 cm long, 15.3 cm wide, and 7.9 cm high, for a total of 300 spores. The spores were covered with a transparent plastic lid to maintain moisture and incubated indoors. Each treatment was replicated three times. During spore germination, the ambient temperature was set at 25°C, and the lighting conditions were set at 2500 lux for 10 hours of light and 14 hours of darkness. The indoor humidity was controlled at 70%.

[0155] On the 30th day, the number of germinated spores was observed and counted under a microscope. On the 90th day, the number of seedlings at the two-leaf, one-heart stage was observed and recorded. The spore germination rate and seedling acquisition rate were calculated according to the formula in Example 1.

[0156] Step 3: Seedling cultivation

[0157] After a period of growth, when the seedlings have grown to three leaves and a height of 2 cm, they are capable of independent life and can be transplanted. After transplanting, the seedlings should be placed in a greenhouse for cultivation. Water regularly weekly to maintain the soil's water holding capacity at 60%, but avoid stagnant water to prevent root rot.

[0158] The formula of the transplanting substrate was the same as that in step 1. Each treatment group was repeated for 100 plants. The growth status of the seedlings was recorded within one month, and the average value of each index was taken.

[0159] The temperature of the greenhouse was controlled at 25°C and the humidity was 70%.

[0160] The light transmittance of the greenhouse is controlled at 50%, and the greenhouse provides a diffuse light environment.

[0161] In order to explore the effects of other breeding conditions on the artificial breeding efficiency of golden retrievers, we set up Example 1 and Examples 7 to 9.

[0162] Example 7

[0163] A method for artificially breeding golden retrievers using spores, comprising:

[0164] Step 1, Matrix Preparation

[0165] The matrix formula is as follows, in parts by weight: 100 parts of garden soil, 20 parts of sawdust, 10 parts of perlite, 10 parts of plant ash, 0.05 parts of manganese sulfate, and 0.3 parts of sodium selenite. The raw materials are thoroughly mixed to obtain the matrix.

[0166] Before use, sterilize the substrate, for example, at 121°C for 20 minutes under high-temperature and high-pressure conditions to completely kill pathogens, insect eggs, and other harmful microorganisms in the soil, creating a sterile environment ideal for spore germination. After sterilization, place the substrate in a nutrient cup, water thoroughly, and set aside.

[0167] Step 2, spore germination

[0168] Golden dog spores were rinsed three times with sterile water to remove dust and pathogens. After rinsing, the spores were evenly spread on the surface of the substrate in nutrient cups. Each nutrient cup was inoculated with 50 spores. The nutrient cups were 22.3 cm long, 15.3 cm wide, and 7.9 cm high, for a total of 300 spores. The spores were covered with a transparent plastic lid to maintain moisture and incubated indoors. Each treatment was replicated three times. During spore germination, the ambient temperature was set at 24°C, and the lighting conditions were set at 2700 lux for 11 hours of light and 13 hours of darkness. The indoor humidity was controlled at 75%.

[0169] On the 30th day, the number of germinated spores was observed and counted under a microscope. On the 90th day, the number of seedlings at the two-leaf, one-heart stage was observed and recorded. The spore germination rate and seedling acquisition rate were calculated according to the formula in Example 1.

[0170] Step 3: Seedling cultivation

[0171] After a period of growth, when the seedlings have grown to three leaves and a height of 2 cm, they are capable of independent life and can be transplanted. After transplanting, the seedlings should be placed in a greenhouse for cultivation. Water regularly weekly to maintain the soil's water holding capacity at 60%, but avoid stagnant water to prevent root rot.

[0172] The formula of the transplanting substrate was the same as that in step 1. Each treatment group was repeated for 100 plants. The growth status of the seedlings was recorded within one month, and the average value of each index was taken.

[0173] The temperature of the greenhouse was controlled at 24°C and the humidity was 80%.

[0174] The light transmittance of the greenhouse is controlled at 50%, and the greenhouse provides a diffuse light environment.

[0175] Example 8

[0176] A method for artificially breeding golden retrievers using spores, comprising:

[0177] Step 1, Matrix Preparation

[0178] The matrix formula is as follows, in parts by weight: 100 parts of garden soil, 20 parts of sawdust, 10 parts of perlite, 10 parts of plant ash, 0.05 parts of manganese sulfate, and 0.3 parts of sodium selenite. The raw materials are thoroughly mixed to obtain the matrix.

[0179] Before use, sterilize the substrate, for example, at 121°C for 20 minutes under high-temperature and high-pressure conditions to completely kill pathogens, insect eggs, and other harmful microorganisms in the soil, creating a sterile environment ideal for spore germination. After sterilization, place the substrate in a nutrient cup, water thoroughly, and set aside.

[0180] Step 2, spore germination

[0181] Golden dog spores were rinsed three times with sterile water to remove dust and pathogens. After rinsing, the spores were evenly spread on the surface of the substrate in nutrient cups. Each nutrient cup was inoculated with 50 spores. The nutrient cups were 22.3 cm long, 15.3 cm wide, and 7.9 cm high, for a total of 300 spores. The spores were covered with a transparent plastic lid to maintain moisture and incubated indoors. Each treatment was replicated three times. During spore germination, the ambient temperature was set at 26°C, and the lighting conditions were set at 3000 lux for 12 hours of light and 12 hours of darkness. The indoor humidity was controlled at 75%.

[0182] On the 30th day, the number of germinated spores was observed and counted under a microscope. On the 90th day, the number of seedlings at the two-leaf, one-heart stage was observed and recorded. The spore germination rate and seedling acquisition rate were calculated according to the formula in Example 1.

[0183] Step 3: Seedling cultivation

[0184] After a period of growth, when the seedlings have grown to three leaves and a height of 2 cm, they are capable of independent life and can be transplanted. After transplanting, the seedlings should be placed in a greenhouse for cultivation. Water regularly weekly to maintain the soil's water holding capacity at 65%, but avoid stagnant water to prevent root rot.

[0185] The formula of the transplanting substrate was the same as that in step 1. Each treatment group was repeated for 100 plants. The growth status of the seedlings was recorded within one month, and the average value of each index was taken.

[0186] The temperature of the greenhouse was controlled at 26°C and the humidity was 75%.

[0187] The light transmittance of the greenhouse is controlled at 50%, and the greenhouse provides a diffuse light environment.

[0188] Example 9

[0189] Based on Example 1, it also includes understory cultivation.

[0190] The method of this embodiment is as follows:

[0191] A method for artificially breeding golden retrievers using spores, comprising:

[0192] Step 1, Matrix Preparation

[0193] The matrix formula is as follows, in parts by weight: 100 parts of garden soil, 20 parts of sawdust, 10 parts of perlite, 10 parts of plant ash, 0.05 parts of manganese sulfate, and 0.3 parts of sodium selenite. The raw materials are thoroughly mixed to obtain the matrix.

[0194] Before use, sterilize the substrate, for example, at 121°C for 20 minutes under high-temperature and high-pressure conditions to completely kill pathogens, insect eggs, and other harmful microorganisms in the soil, creating a sterile environment ideal for spore germination. After sterilization, place the substrate in a nutrient cup, water thoroughly, and set aside.

[0195] Step 2, spore germination

[0196] Golden dog spores were rinsed three times with sterile water to remove dust and pathogens. After rinsing, the spores were evenly spread on the surface of the substrate in nutrient cups. Each nutrient cup was inoculated with 50 spores. The nutrient cups were 22.3 cm long, 15.3 cm wide, and 7.9 cm high, for a total of 300 spores. When spores germinated, they were covered with a transparent plastic cover to maintain moisture. The spores were then incubated indoors under low temperature. Each treatment was repeated three times. The ambient temperature was set at 25°C, and the lighting conditions were set at 2500 lux for 10 hours of light and 14 hours of darkness. The indoor humidity was controlled at 70%.

[0197] On the 30th day, the number of germinated spores was observed and counted under a microscope. On the 90th day, the number of seedlings at the two-leaf, one-heart stage was observed and recorded. The spore germination rate and seedling acquisition rate were calculated according to the formula in Example 1.

[0198] Step 3: Seedling cultivation

[0199] After a period of growth, when the seedlings have grown to three leaves and a height of 2 cm, they are capable of independent life and can be transplanted. After transplanting, the seedlings should be placed in a greenhouse for cultivation. Water regularly weekly to maintain the soil's water holding capacity at 60%, but avoid stagnant water to prevent root rot.

[0200] The formula of the transplanting substrate was the same as that in step 1. Each treatment group was repeated for 100 plants. The growth status of the seedlings was recorded within one month, and the average value of each index was taken.

[0201] The temperature of the greenhouse was controlled at 25°C and the humidity was 70%.

[0202] The light transmittance of the greenhouse is controlled at 50%, and the greenhouse provides a diffuse light environment.

[0203] Step 4: After the seedling cultivation is completed, carry out under-forest cultivation

[0204] The selection of a forest cultivation site is crucial to the subsequent growth of golden retrievers. After research and screening, Chinese fir forests in subtropical areas with an annual average temperature above 5°C have become ideal forest cultivation sites.

[0205] Covering the soil under the forest with mulch can increase soil temperature, retain soil moisture, and promote rapid root growth. Then, transplant the seedlings and build a sunshade over them to provide them with a suitable diffused light environment. After two months of growth, the seedlings gradually adapt to the forest environment. At this time, remove the mulch and sunshade, allowing the seedlings to continue growing and developing in the natural forest environment, making full use of the rich natural resources under the forest to promote their healthy growth.

[0206] In order to more clearly illustrate the differences between the above embodiments and the comparison, the present invention provides Table 1 and Table 2.

[0207] Table 1 Differences in matrix formulations (in dry weight parts)

[0208] Grouping Farmland soil sawdust Perlite plant ash Manganese sulfate Sodium selenite Example 1 100 20 10 10 0.05 0.3 Example 2 100 20 10 10 0.1 0.3 Example 3 100 20 10 10 0.15 0.3 Example 4 100 20 10 10 0.1 0.4 Example 5 100 20 10 10 0.1 0.5 Example 6 100 25 15 12 0.1 0.4 Blank control 100 20 10 10 0 0 Single factor control 1 100 20 10 10 0.05 0 Single factor control 2 100 20 10 10 0 0.3

[0209] For other differences in breeding conditions, see Table 2.

[0210] Table 2 Other breeding conditions

[0211]

[0212] The experimental results are shown in Table 3.

[0213] Table 3 Experimental results

[0214]

[0215]

[0216] The results of Table 3 are analyzed as follows:

[0217] (1) Spore germination rate

[0218] Using the method of the present invention, the spore germination rate and seedling yield rate were significantly improved. The spore germination rate of the blank control was only 33%, and the seedling yield rate was only 10%. The spore germination rate and seedling yield rate of the single factor control 1 and single factor control 2 were also significantly lower than those of the examples.

[0219] The spore germination rate directly reflects the natural renewal ability and potential development trend of golden dogs. A higher spore germination rate means that more seeds can successfully break through the dormant state. From the perspective of the ecosystem, a high spore germination rate can ensure the proportion of golden dogs in the community and maintain the species diversity and structural stability of the ecosystem. For artificial breeding of golden dogs, the spore germination rate is even more important. Artificial breeding of golden dogs aims to expand the population size for ecological restoration or protection research. By increasing the spore germination rate, the breeding cost can be reduced and the breeding efficiency can be improved. A high spore germination rate can not only quickly increase the number of golden dogs, but also provide rich materials for subsequent genetic diversity research, which helps to cultivate varieties that are more adaptable to environmental changes. Therefore, the effect of improving the spore germination rate of the present invention has great ecological significance.

[0220] (2) Seedling acquisition rate and seedling survival rate after transplanting

[0221] The method of the embodiment of the present invention can maintain a seedling acquisition rate of over 70%, while the control group only had 10%. The survival rate of the seedlings after transplanting in the blank control group was 22%, while the method of the embodiment of the present invention can stably maintain the survival rate of the seedlings after transplanting at 83% to 85%, greatly improving the conversion rate of breeding results.

[0222] The seedling acquisition rate is highly correlated with material utilization and work efficiency. The method of the embodiment of the present invention can improve the seedling acquisition rate, which means that a large amount of cost and time cost can be saved, and material utilization and work efficiency can be improved.

[0223] The survival rate of seedlings after transplanting refers to the proportion of seedlings that survive and grow to a certain stage after germination. It is a key link in the growth process of golden dog fern, directly affecting the transition of the population from the seedling stage to the adult stage. From the perspective of ecological balance, the survival rate of seedlings after transplanting affects the energy flow and material circulation of the entire ecosystem. As a producer in the ecosystem, the survival of golden dog fern seedlings means that more photosynthetic products are fixed, providing food and habitat for other organisms. In terms of artificial cultivation, improving the survival rate of seedlings after transplanting is the key to ensuring cultivation results. A high survival rate of seedlings after transplanting can reduce resource waste in the cultivation process, improve the economic and ecological benefits of artificial propagation, and accelerate the rejuvenation and population expansion of golden dog fern.

[0224] (3) Growth uniformity

[0225] This method produces seedlings of uniform physiological age and uniform growth. The coefficient of variation for all seedling growth indicators was kept within 10%. This demonstrates that this propagation technique effectively ensures consistent seedling growth, providing a good foundation for large-scale seedling production and cultivation, facilitating unified management and maintenance, and improving production efficiency.

[0226] Growth uniformity refers to the degree of consistency in the growth of individuals within a population of golden dog fern. In natural environments, golden dog fern populations with high growth uniformity are more competitive. Uniform growth of individuals optimizes environmental resources and reduces competition among individuals. In artificial cultivation and management, growth uniformity facilitates unified cultivation and harvesting, saving manpower and material resources, and improving harvesting efficiency and product quality.

[0227] (4) Seedling development

[0228] The present invention evaluates the development of seedlings based on the number of new leaves produced, leaf length and leaf width. The results show that compared with the blank control, single-factor control 1 and single-factor control 2, the number of new leaves produced, leaf length and leaf width are significantly improved by using the embodiment method of the present invention.

[0229] It should be noted that when numerical ranges are involved in the present invention, it should be understood that both endpoints of each numerical range and any value between the two endpoints can be selected. Since the steps and methods used are the same as those in the embodiments, in order to avoid redundancy, the present invention describes preferred embodiments. Although preferred embodiments of the present invention have been described, those skilled in the art may make additional changes and modifications to these embodiments once they understand the inventive concept of the present invention, and such changes and modifications fall within the scope of the present invention.

[0230] Obviously, those skilled in the art may make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. If these modifications and variations of the present invention fall within the scope of the equivalent technology of the present invention, the present invention is intended to include these modifications and variations.

Claims

1. A method for artificially breeding golden retrievers using spores, characterized in that: include: The matrix formula is as follows in parts by weight: 100 parts of garden soil, 20 to 25 parts of sawdust, 10 to 15 parts of perlite, 10 to 12 parts of plant ash, 0.05 to 0.15 parts of manganese sulfate, and 0.3 to 0.5 parts of sodium selenite; Spores germinate, and the environmental temperature for spore germination is 24℃~26℃; The germinated seedlings are transplanted into greenhouses for cultivation.

2. The method for artificially breeding golden retrievers using spores according to claim 1, characterized in that: The matrix formula is as follows: 100 parts of garden soil, 20 parts of sawdust, 10 parts of perlite, 10 parts of wood ash, 0.1 parts of manganese sulfate, and 0.4 parts of sodium selenite.

3. The method for artificially breeding golden retrievers using spores according to claim 1, characterized in that: The sawdust is poplar sawdust powder.

4. The method for artificially breeding golden retrievers using spores according to claim 3, characterized in that: Spore germination was carried out indoors, and the lighting conditions were set to: 2500 lux ~ 3000 lux for 10h ~ 12h.

5. The method for artificially breeding golden retrievers using spores according to claim 4, characterized in that: The indoor humidity is set to 70% to 80%.

6. The method for artificially breeding golden retrievers using spores according to claim 1, characterized in that: The temperature in the greenhouse is 24℃~26℃, and the humidity is 70%~75%.

7. The method for artificially breeding golden retrievers using spores according to claim 6, characterized in that: Maintain soil water holding capacity at 60% to 65%.

8. A method for improving the artificial breeding efficiency of golden retrievers, characterized in that: The matrix is ​​the matrix according to claim 1.

9. The use according to claim 8, characterized in that The improvement of the artificial breeding efficiency of golden dog refers to improving the spore germination rate, the seedling acquisition rate, the survival rate of the seedlings after transplanting, and the promotion of seedling development.

10. The use according to claim 9, characterized in that The promoting of seedling development refers to accelerating the production of new leaves and increasing leaf length and leaf width.

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