Standardized breeding and cultivating method for genuine medicinal material seeds

Abstract

The invention relates to the technical field of medicinal material cultivation, and discloses a genuine medicinal material seed standardized breeding cultivation method which comprises the following steps: selecting excellent plants in a traditional high-quality producing area during seed selection, and performing gene detection on provenance plants to screen excellent gene labeled plants; the seeds are washed by clear water, disinfected by potassium permanganate and soaked by borax; constant-temperature and constant-humidity box variable-temperature treatment is adopted for accelerating germination, and ethylene gas is added; a specific matrix formula and temperature, humidity and illumination control are used for seedling raising, and a microbial agent is added; dipping a rooting powder solution during transplanting; performing field management according to fertility topdressing and water control; the pest control adopts a physical, biological and chemical combination mode. The method ensures excellent genetic quality of seeds, improves the survival rate and growth vigor of seedlings, improves the content, yield and quality of effective components of medicinal materials, reduces pesticide residues, enhances market competitiveness, reduces cost, brings good economic benefits to planters, and promotes development of genuine medicinal material industry.

Description

Technical Field

[0001] The invention relates to the technical field of medicinal material cultivation, and in particular to a standardized selection and cultivation method for authentic medicinal material seeds. Background Art

[0002] Authentic medicinal materials refer to Chinese medicinal materials that grow in specific natural environments and regions, and have been verified by long-term practice to have unique qualities, excellent medicinal effects and stable quality, such as Chuanxiong from Sichuan and Angelica sinensis from Gansu. The active ingredients contained in authentic medicinal materials are richer and purer, which can ensure the reliability and stability of clinical treatment effects. They play a key role in the field of traditional Chinese medicine and are an important material basis for the precise treatment of traditional Chinese medicine prescriptions. They provide high-quality medicinal materials for many classic prescriptions, and also carry the profound heritage of traditional Chinese medicine culture. They have irreplaceable value in promoting the healthy development of the traditional Chinese medicine industry, enhancing international competitiveness, and inheriting and innovating the cause of traditional Chinese medicine.

[0003] There are many drawbacks in seed selection and cultivation of traditional authentic medicinal materials. Seed selection relies on experience, lacks precise screening, and is difficult to ensure excellent genetic characteristics. Seed treatment and germination techniques are simple, which affects germination and initial growth. There is no standard for seedling medium, seedlings grow poorly, are susceptible to diseases, and have low survival rates. Field management, fertilization and watering are difficult to be precise, resulting in fertilizer waste, soil problems, and drought and flood disasters, affecting the growth and yield of medicinal materials. Pest and disease control relies on chemical pesticides, which cause drug resistance and residues, pollute the environment, reduce quality, and threaten health. There is also a lack of scientific knowledge of ecology and growth laws, which restricts industrial development. Therefore, there is an urgent need for a standardized and scientific method for seed selection and cultivation of authentic medicinal materials to solve the above problems, improve the quality and yield of authentic medicinal materials, and ensure their quality safety and the healthy development of the industry. Summary of the invention

[0004] In view of the deficiencies in the prior art, the present invention provides a standardized method for selecting and breeding authentic medicinal material seeds, which solves the problem of imperfect seed selection and breeding in traditional authentic medicinal material planting.

[0005] To achieve the above objectives, the present invention is implemented through the following technical solutions: a standardized breeding and cultivation method for authentic medicinal material seeds, comprising the following steps: S1. Seed selection In the traditional high-quality production areas of authentic medicinal materials, we select plants with excellent traits and stable yields for more than two consecutive years as seed sources, and collect fully mature, plump, disease-free and insect-free seeds during the seed maturity season; S2. Seed treatment Rinse the collected seeds with clean water for 10-15 minutes, then soak them in 0.5%-1% potassium permanganate solution for 10-20 minutes for disinfection, then rinse them with clean water and place them in a ventilated and cool place to dry, so that the moisture content of the seeds is maintained at 8%-12%; S3. Germination Place the sterilized and dried seeds in a constant temperature and humidity chamber at 20℃-25℃ and 60%-70% humidity, and use variable temperature treatment, treat at 15℃-20℃ for 8-10 hours a day, and at 25℃-30℃ for 6-8 hours, for 5-7 days. During this period, rinse the seeds with clean water 1-2 times a day until the seed whitening rate reaches 30%-40%; S4. Seedling cultivation: Sow the whitened seeds on the seedling medium, which is a mixture of leaf mold, perlite and vermiculite in a volume ratio of 3:1:1. The sowing depth is 1-2 cm. Maintain the seedling environment temperature at 20℃-25℃, humidity at 60%-70%, light intensity at 2000-3000 lux, and light time at 12-14 hours per day. S5. Transplanting When the seedlings grow to 4-6 true leaves, choose a cloudy day or evening for transplanting. Harden the seedlings for 3-5 days before transplanting, and transplant them to a planting field that has been 30-40 cm deep plowed, 2000-3000 kg / mu of organic fertilizer and 50-100 kg / mu of superphosphate applied. The spacing between transplanted plants is 20-30 cm × 30-40 cm. Water the roots in time after transplanting. S6. Field management Weed and loosen the soil regularly. During the growing period, apply 10-15 kg / mu of nitrogen fertilizer and 20-30 kg / mu of compound fertilizer according to the soil fertility. Water in time during drought to keep the soil moisture content at 40%-60%. Drain water in time to prevent waterlogging in the rainy season. S7. Pest and disease control Use a combination of physical, biological and chemical control methods to control pests and diseases; S8. Seed collection The authentic medicinal materials are harvested when their fruits are ripe and their seeds are fully mature. After harvesting, the fruits are dried in the sun for 3-5 days, and then threshed to obtain the seeds. The seeds are dried at 30℃-40℃ to a moisture content of 6%-8%, packaged in sealed bags and stored in a cool and dry place.

[0006] Preferably, in the seed selection step, genetic testing is performed on the selected source plants to screen out plants with specific excellent gene markers, and the specific excellent gene markers are related to the active ingredient content and stress resistance of the medicinal material.

[0007] Preferably, the gene detection comprises the following steps: S1.1. Collect fresh leaves of provenance plants, extract plant genomic DNA using the CTAB method, and use a nucleic acid protein detector to measure the purity and concentration of DNA to ensure that the OD260 / OD280 value of DNA is between 1.8-2.0 and the concentration is not less than 50ng / μL. Dilute qualified DNA samples to a working concentration of 20-30ng / μL for later use; S1.2. Design specific primers for the key enzyme genes in the biosynthesis pathway of the active ingredients of the authentic medicinal materials and the transcription factor gene fragments related to stress resistance. The primer sequences are designed by PrimerPremier5.0 software, with an annealing temperature between 55°C and 65°C, a primer length of 18-25bp, and a GC content between 40% and 60%. After the primers are synthesized, they are purified by PAGE; S1.3. Real-time fluorescence quantitative PCR technology was used to quantitatively analyze the above genes. The reaction system was 20 μL, including 10 μL of 2×SYBR Green PCR Master Mix, 0.4 μL of upstream and downstream primers, 2 μL of template DNA and 7.2 μL of ddH2O. The PCR reaction program was: 95℃ pre-denaturation for 5 min; 95℃ denaturation for 15 s, 55℃-65℃ annealing for 30 s, 72℃ extension for 30 s, a total of 40 cycles; finally, melting curve analysis was performed to determine the specificity of the amplified product; S1.4. Based on the results of real-time fluorescence quantitative PCR, plants with relative expression levels of target genes that are more than 30% higher than the average level of the same production area are screened as seed sources with specific excellent gene markers. The target genes include but are not limited to key enzyme genes that control the content of effective ingredients and transcription factor genes that participate in stress resistance response.

[0008] Preferably, in the seed treatment step, after soaking in potassium permanganate solution, the seeds are further soaked in 0.1%-0.2% borax solution for 5-10 minutes.

[0009] Preferably, in the germination step, an appropriate amount of ethylene gas is added in a constant temperature and humidity chamber, and the ethylene gas concentration is 10-20 ppm to promote seed germination and seedling growth.

[0010] Preferably, in the seedling raising step, a microbial agent is added to the seedling raising matrix, and the added amount of the microbial agent is 0.5%-1% of the weight of the matrix. The microbial agent includes beneficial bacteria such as Bacillus subtilis and Trichoderma harzianum.

[0011] Preferably, in the transplanting step, the roots of the seedlings are dipped in a rooting powder solution during transplanting, and the concentration of the rooting powder solution is 0.1%-0.2%.

[0012] Preferably, in the field management step, during topdressing, a fertilizer solution is evenly applied to the roots of the plants by drip irrigation or sprinkler irrigation, and the concentration of the fertilizer solution is 0.5%-1%.

[0013] Preferably, in the pest control step, physical control uses insect-proof nets and yellow boards to lure and kill pests, and biological control releases natural enemies of pests. When the pests and diseases occur seriously, high-efficiency, low-toxicity, and low-residue pesticides are used for control.

[0014] The present invention provides a standardized method for selecting and breeding authentic medicinal material seeds. It has the following beneficial effects: 1. The present invention ensures that the selected seeds have high genetic quality through strict seed selection steps, including selecting excellent plants in traditional high-quality production areas and performing genetic testing on the source plants to screen plants with specific excellent gene markers. The genes related to the active ingredient content and stress resistance of the medicinal materials are optimized, thereby laying the foundation for the subsequent cultivation of high-quality authentic medicinal materials.

[0015] 2. During the seedling raising and transplanting process, the present invention adopts scientific and reasonable seedling raising substrate formula, temperature, humidity and light control, as well as measures such as adding microbial agents and rooting powder solution, which significantly improves the survival rate and growth potential of seedlings, makes the seedlings stronger and more adaptable to the environment, reduces the loss of seedlings, reduces production costs, and improves planting benefits.

[0016] 3. The present invention uses refined operations in seed treatment, germination and field management. The combination of potassium permanganate and borax for seed treatment, temperature change and ethylene addition for germination, precise topdressing according to fertility in the field and reasonable water control can promote plant growth, enhance stress resistance and disease resistance, increase the content and yield of effective ingredients in medicinal materials, and adopt a combination of physical, biological and chemical pest control methods. While controlling pests, the amount of pesticides used is reduced, residues are reduced, the quality and safety of medicinal materials are guaranteed, the market demand for high quality is met, competitiveness is enhanced, and the economic benefits of growers are increased. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 The present invention is a flow chart of the method. DETAILED DESCRIPTION

[0018] The following will be combined with the drawings in the specification of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0019] Embodiment 1: Please refer to the attached Figure 1The embodiment of the present invention provides a standardized method for selecting and breeding authentic medicinal material seeds, comprising the following steps: S1. Seed selection: In Hunyuan, Shanxi, a traditional high-quality production area of ​​Astragalus, 50 plants with thick stems, lush leaves, stable yields for more than two consecutive years, and high astragaloside content were selected as provenance. In the seed maturity season, fully mature, plump, and pest-free seeds were collected. Genetic testing was performed on the selected provenance plants, and fresh leaves were collected. Plant genomic DNA was extracted using the CTAB method. The OD260 / OD280 value was 1.9 and the concentration of DNA was 60 ng / μL after determination by a nucleic acid protein detector. The DNA was diluted to the working The concentration was 25 ng / μL for standby use. Specific primers were designed for the key enzyme genes in the astragaloside synthesis pathway and the transcription factor gene fragments related to stress resistance. The primer annealing temperature was 60°C, the length was 20 bp, and the GC content was 50%. After synthesis, it was purified by PAGE and the gene was quantitatively analyzed by real-time fluorescence quantitative PCR technology. The reaction system was 20 μL, containing 10 μL of 2×SYBR Green PCR Master Mix, 0.4 μL of upstream and downstream primers, 2 μL of template DNA and 7.2 μL of ddH 2 O, PCR program is 95℃ pre-denaturation for 5min; 95℃ denaturation for 15s, 60℃ annealing for 30s, 72℃ extension for 30s, a total of 40 cycles, melting curve analysis to determine the specificity of the amplified product, screened out 20 plants with a relative expression of the target gene 35% higher than the average level of the same production area as the provenance, ensuring the excellent genetic characteristics of the seeds. Compared with traditional random selection or selection based only on appearance traits, the accuracy of seed selection is greatly improved. The provenance selected by genetic testing has more advantages in astragaloside content and stress resistance, which is expected to improve the quality and yield stability of medicinal materials, reduce the risk of quality differences and yield fluctuations caused by genetic factors, and provide reliable starting materials for the cultivation of high-quality astragalus. S2. Seed treatment: Rinse the collected seeds with clean water for 12 minutes, then soak them in 0.8% potassium permanganate solution for 15 minutes for disinfection, then rinse them with clean water, place them in a ventilated and cool place to dry, keep the moisture content of the seeds at 10%, and then soak them in 0.15% borax solution for 8 minutes. Potassium permanganate has strong oxidizing properties and can effectively kill a variety of pathogens. It destroys the cell structure and biological activity of pathogens through oxidation, disinfects the seeds, and prevents the seeds from carrying pathogens and causing diseases in the subsequent cultivation process. Borax solution soaking further optimizes the physiological state of the seeds, so that the seeds can start the germination process faster in the subsequent germination and seedling raising process, grow stronger, enhance the resistance of seedlings to adverse environments, reduce cultivation failures and poor growth caused by seed problems themselves, and improve cultivation efficiency and success rate; S3. Germination: The sterilized and dried seeds were placed in a constant temperature and humidity chamber at a temperature of 22°C and a humidity of 65%. The seeds were treated at 18°C ​​for 9 hours and 28°C for 7 hours every day for 6 days. During this period, the seeds were rinsed with clean water twice a day until the seed whitening rate reached 35%. During the germination process, ethylene gas was added to the constant temperature and humidity chamber at a concentration of 15ppm. The precise temperature and humidity control and variable temperature treatment combined with the addition of ethylene gas significantly improved the germination effect of the seeds. Compared with the traditional single temperature germination method, this variable temperature treatment can more effectively break the seed dormancy, shorten the seed germination time, and make the seed whitening rate reach 35%, which improves the germination speed and uniformity of the seeds, and is conducive to the subsequent seedling management and synchronous growth of seedlings. S4. Seedling cultivation: sow the whitened seeds on a seedling cultivation substrate mixed with leaf mold, perlite and vermiculite in a volume ratio of 3:1:1, with a sowing depth of 1.5 cm, and maintain the seedling cultivation environment temperature at 23°C, humidity at 65%, light intensity at 2500 lux, and light time for 13 hours per day. Add microbial agents to the seedling cultivation substrate in an amount of 0.8% of the substrate weight, including Bacillus subtilis and Trichoderma harzianum. Bacillus subtilis can produce a variety of antibiotics and enzyme substances, inhibit the growth and reproduction of pathogens in the soil, and protect the seedling roots from pathogens. Trichoderma harzianum can decompose organic matter in the soil, release nutrients, and promote the absorption and utilization of nutrients by the roots. At the same time, its mycelium can also wrap around the roots to enhance the root resistance and fixation ability to the soil; S5. Transplanting: When the seedlings grow to 5 true leaves, choose a cloudy day for transplanting. Before transplanting, harden the seedlings for 4 days and transplant them to a planting field that has been 35 cm deep plowed, 2500 kg / mu of organic fertilizer and 80 kg / mu of superphosphate applied. The transplanting row spacing is 25 cm × 35 cm. When transplanting, dip the roots of the seedlings in a 0.15% rooting powder solution. After transplanting, water the roots in time. Dipping the roots in the rooting powder solution and watering the roots in time further promote the growth and recovery of the root system, improve the survival rate and growth vitality of the seedlings, and ensure the smooth progress of the transplanting work and the early healthy growth of the Astragalus plants. S6. Field management: Regularly weed and loosen the soil. During the growth period, apply nitrogen fertilizer 12 kg / mu and compound fertilizer 25 kg / mu according to the soil fertility. Water in time during drought to keep the soil moisture content at 50%. Drain in time to prevent waterlogging during the rainy season. When applying fertilizer, use drip irrigation to evenly apply the fertilizer solution to the roots of the plants. The concentration of the fertilizer solution is 0.8%; S7. Pest and disease control: Physical control uses insect-proof nets and yellow boards to lure and kill pests, and biological control releases natural enemies of pests such as seven-spotted ladybugs. When pests and diseases are serious, use 10% highly effective chlorflucythrinate emulsifiable concentrate diluted 2000 times to spray for pest control, and use 75% thiophanate-methyl wettable powder diluted 1000 times to spray for disease control. Highly effective chlorflucythrinate is a highly effective, low-toxic, low-residue insecticide that can quickly kill pests through contact and stomach poisoning. Its diluted low-concentration solution can not only ensure the control effect on pests, but also reduce the harm to the environment and beneficial organisms. Use 75% thiophanate-methyl wettable powder diluted 1000 times to spray for disease control, which can inhibit the growth and reproduction of various pathogens, and prevent the invasion and spread of pathogens by forming a protective film on the surface of the plant, thus protecting the plant from the invasion of diseases; S8. Seed harvesting: When the astragalus fruit is ripe and the seeds are fully mature, the fruit is harvested. After harvesting, the fruit is dried in the sun for 4 days, and then threshed to obtain seeds. The seeds are dried at 35°C to a moisture content of 7%, and then packaged in a sealed bag and stored in a cool and dry place. According to tests, the germination rate of astragalus seeds cultivated by this method is 20% higher than that of the traditional method, the astragaloside content is increased by 15%, and the seedling survival rate is increased by 25%.

[0020] Embodiment 2: The embodiment of the present invention provides a standardized method for selecting and breeding authentic medicinal material seeds, comprising the following steps: S1. Seed selection: In Ningxia Zhongning, a traditional high-quality production area of ​​wolfberry, 30 plants with large fruits, bright colors, stable yields for more than two consecutive years, and high content of wolfberry polysaccharides were selected as provenance. After collecting mature seeds, genetic testing was performed on the provenance plants. The extracted DNA was measured to have an OD260 / OD280 value of 1.85 and a concentration of 55 ng / μL. The DNA was diluted to 22 ng / μL for standby use. The designed primers had an annealing temperature of 58°C, a length of 22 bp, and a GC content of 45%. After PAGE purification, they were used for real-time fluorescence quantitative PCR. The reaction system and procedure were the same as in Example 1. 15 plants with a relative expression level of the target gene that was 40% higher than the average level in the same production area were screened out as provenance; S2. Seed treatment: Rinse the seeds with clean water for 10 minutes, soak them in 0.6% potassium permanganate solution for 12 minutes for disinfection, wash and dry them to a moisture content of 9%, and then soak them in 0.1% borax solution for 6 minutes; S3. Germination: In a constant temperature and humidity box at 20℃ and 62% humidity, the temperature was changed to 16℃ for 8 hours and 26℃ for 7 hours for 5 days. The seeds were rinsed once a day and the whitening rate reached 32%. 12ppm ethylene gas was added into the box; S4. Seedling cultivation: Sow the white seeds in a 3:1:1 mixture of leaf mold, perlite and vermiculite, 1 cm deep, 20°C, 62% humidity, 2200 lux, 12 hours of light. Add 0.6% microbial agent containing Bacillus subtilis to the substrate; S5. Transplanting: Transplant on a cloudy day when the seedlings have 4 true leaves, harden the seedlings for 3 days, and move to a field with a 32 cm deep tillage, 2200 kg / mu of organic fertilizer and 60 kg / mu of superphosphate, with a row spacing of 22 cm × 32 cm. Dip the seedlings in 0.12% rooting powder solution and water them for rooting. S6. Field management: regular weeding, soil loosening, topdressing of nitrogen fertilizer 10 kg / mu, compound fertilizer 22 kg / mu, watering to 45% moisture content during drought, drainage in rainy season. Drip irrigation fertilization concentration 0.6%; S7 pest control: Use insect nets and yellow boards for physical control, release predatory mites for biological control, and use 2500 times diluted 8% highly effective cyhalothrin emulsifiable concentrate and 800 times diluted 65% mancozeb wettable powder for control when the disease is serious; S8 Seed Harvest: After the fruit matures, it is sun-dried for 3 days and threshed, then dried at 32℃ until the moisture content reaches 6.5%, and bagged and stored in a cool place. The germination rate of wolfberry seeds cultivated by this method is increased by 18%, the content of wolfberry polysaccharides is increased by 12%, and the survival rate of seedlings is increased by 22%.

[0021] Embodiment three: The embodiment of the present invention provides a standardized method for selecting and breeding authentic medicinal material seeds, comprising the following steps: S1. Seed selection: In Pingyi, Shandong, a traditional high-quality production area of ​​honeysuckle, 40 plants with luxuriant flowers and leaves, high chlorogenic acid content and stable yield were selected as provenance. After seed collection, genetic testing was performed to obtain DNA with an OD260 / OD280 value of 1.92 and a concentration of 58 ng / μL, which was diluted to 28 ng / μL. The primer annealing temperature was 62°C, the length was 19 bp, and the GC content was 55%. It was used for quantitative PCR to screen out 20 provenances with a 38% higher expression of the target gene; S2. Seed treatment: wash with water for 14 minutes, soak in 0.9% potassium permanganate for 18 minutes, dry to 11% moisture content, and then soak in 0.2% borax for 9 minutes; S3. Germination: 24°C, 68% humidity constant temperature and humidity chamber, 19°C for 9 hours, 29°C for 7 hours, 6 days, rinse twice a day until the white rate reaches 38%, add 18ppm ethylene gas; S4. Seedling cultivation: sow in a 3:1:1 substrate, 1.8 cm deep, 24°C, 68% humidity, 2800 lux light intensity, 14 hours of light, and add 0.9% of the inoculant containing Trichoderma harzianum; S5. Transplanting: Transplant on a cloudy day when 6 true leaves have grown, harden the seedlings for 5 days, move to a field with a 38 cm deep tillage, 2800 kg / mu of organic fertilizer and 90 kg / mu of superphosphate, and a row spacing of 28 cm × 38 cm. Dip in 0.18% rooting powder solution and water for rooting. S6. Field management: weeding, loosening the soil, topdressing nitrogen 14 kg / mu, compound fertilizer 28 kg / mu, watering to 55% moisture content during drought, drainage in rainy season, sprinkler irrigation 0.9% concentration of fertilizer; S7. Pest and disease control: Physically use yellow insect-proof nets and boards, biologically release parasitic bees, and for severe cases, use 12% highly effective cyhalothrin emulsifiable concentrate 1800 times diluted and 80% carbendazim wettable powder 600 times diluted for control; S8 seed harvesting: After ripening, dry them in the sun for 5 days to thresh them, bake them at 38℃ until the moisture content reaches 7.5% and store them in a cool place. This method can increase the germination rate of honeysuckle seeds by 22%, the chlorogenic acid content by 18%, and the survival rate of seedlings by 30%.

[0022] In summary, the standardized selection and breeding method for authentic medicinal material seeds of the present invention has made remarkable progress in many aspects. By selecting seeds in traditional high-quality production areas and combining genetic testing, excellent seed sources are accurately screened to lay the foundation for high-quality medicinal materials. The refined operations and optimized technologies in seed treatment, germination, seedling cultivation, transplanting and field management have significantly improved the seed germination rate, seedling survival rate and the content of effective ingredients in medicinal materials. In addition, comprehensive means are adopted for disease and pest control to reduce pesticide residues, which has effectively promoted the high-quality and efficient production of authentic medicinal materials.

[0023] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A standardized method for selecting and breeding authentic medicinal material seeds, characterized in that: The following steps are involved: S1. Seed selection In the traditional high-quality production areas of authentic medicinal materials, we select plants with excellent traits and stable yields for more than two consecutive years as seed sources, and collect fully mature, plump, disease-free and insect-free seeds during the seed maturity season; S2. Seed treatment Rinse the collected seeds with clean water for 10-15 minutes, then soak them in 0.5%-1% potassium permanganate solution for 10-20 minutes for disinfection, then rinse them with clean water and place them in a ventilated and cool place to dry, so that the moisture content of the seeds is maintained at 8%-12%; S3. Germination Place the sterilized and dried seeds in a constant temperature and humidity chamber at 20℃-25℃ and 60%-70% humidity, and use variable temperature treatment, treat at 15℃-20℃ for 8-10 hours a day, and at 25℃-30℃ for 6-8 hours, for 5-7 days. During this period, rinse the seeds with clean water 1-2 times a day until the seed whitening rate reaches 30%-40%; S4. Seedling cultivation: Sow the whitened seeds on the seedling medium, which is a mixture of leaf mold, perlite and vermiculite in a volume ratio of 3:1:

1. The sowing depth is 1-2 cm. Maintain the seedling environment temperature at 20℃-25℃, humidity at 60%-70%, light intensity at 2000-3000 lux, and light time at 12-14 hours per day. S5. Transplanting When the seedlings grow to 4-6 true leaves, choose a cloudy day or evening for transplanting. Harden the seedlings for 3-5 days before transplanting, and transplant them to a planting field that has been 30-40 cm deep plowed, 2000-3000 kg / mu of organic fertilizer and 50-100 kg / mu of superphosphate applied. The spacing between transplanted plants is 20-30 cm × 30-40 cm. Water the roots in time after transplanting. S6. Field management Weed and loosen the soil regularly. During the growing period, apply 10-15 kg / mu of nitrogen fertilizer and 20-30 kg / mu of compound fertilizer according to the soil fertility. Water in time during drought to keep the soil moisture content at 40%-60%. Drain water in time to prevent waterlogging in the rainy season. S7. Pest and disease control Use a combination of physical, biological and chemical control methods to control pests and diseases; S8. Seed collection The authentic medicinal materials are harvested when their fruits are ripe and their seeds are fully mature. After harvesting, the fruits are dried in the sun for 3-5 days, and then threshed to obtain the seeds. The seeds are dried at 30℃-40℃ to a moisture content of 6%-8%, packaged in sealed bags and stored in a cool and dry place.

2. The method for standardized selection and breeding of authentic medicinal material seeds according to claim 1, characterized in that: In the seed selection step, the selected source plants are subjected to gene detection to screen out plants with specific excellent gene markers, wherein the specific excellent gene markers are related to the active ingredient content and stress resistance of the medicinal material.

3. The method for standardized selection and breeding of authentic medicinal material seeds according to claim 2, characterized in that: The genetic testing includes the following steps: S1.

1. Collect fresh leaves of provenance plants, extract plant genomic DNA using the CTAB method, and use a nucleic acid protein detector to measure the purity and concentration of DNA to ensure that the OD260 / OD280 value of DNA is between 1.8-2.0 and the concentration is not less than 50ng / μL. Dilute qualified DNA samples to a working concentration of 20-30ng / μL for later use; S1.

2. Design specific primers for the key enzyme genes in the biosynthesis pathway of the active ingredients of the authentic medicinal materials and the transcription factor gene fragments related to stress resistance. The primer sequences are designed by PrimerPremier5.0 software, with an annealing temperature between 55°C and 65°C, a primer length of 18-25bp, and a GC content between 40% and 60%. After the primers are synthesized, they are purified by PAGE; S1.

3. Real-time fluorescence quantitative PCR technology was used to quantitatively analyze the above genes. The reaction system was 20 μL, including 10 μL of 2×SYBR Green PCR Master Mix, 0.4 μL of upstream and downstream primers, 2 μL of template DNA and 7.2 μL of ddH2O. The PCR reaction program was: 95℃ pre-denaturation for 5 min; 95℃ denaturation for 15 s, 55℃-65℃ annealing for 30 s, 72℃ extension for 30 s, a total of 40 cycles; finally, melting curve analysis was performed to determine the specificity of the amplified product; S1.

4. Based on the results of real-time fluorescence quantitative PCR, plants with relative expression levels of target genes that are more than 30% higher than the average level of the same production area are screened as seed sources with specific excellent gene markers. The target genes include but are not limited to key enzyme genes that control the content of effective ingredients and transcription factor genes that participate in stress resistance response.

4. The method for standardized selection and cultivation of authentic medicinal material seeds according to claim 1, characterized in that: In the seed treatment step, after being soaked in the potassium permanganate solution, the seeds are further soaked in a 0.1%-0.2% borax solution for 5-10 minutes.

5. The method for standardized selection and breeding of authentic medicinal material seeds according to claim 1, characterized in that: In the germination step, a proper amount of ethylene gas is added into the constant temperature and humidity chamber, and the ethylene gas concentration is 10-20 ppm.

6. The method for standardized selection and cultivation of authentic medicinal material seeds according to claim 1, characterized in that: In the seedling raising step, a microbial agent is added to the seedling raising matrix, and the added amount of the microbial agent is 0.5%-1% of the weight of the matrix. The microbial agent includes Bacillus subtilis and Trichoderma harzianum.

7. The method for standardized selection and cultivation of authentic medicinal material seeds according to claim 1, characterized in that: In the transplanting step, the roots of the seedlings are dipped in a rooting powder solution during transplanting, and the concentration of the rooting powder solution is 0.1%-0.2%.

8. The method for standardized selection and breeding of authentic medicinal material seeds according to claim 1, characterized in that: In the field management step, during topdressing, a fertilizer solution is evenly applied to the roots of the plants by drip irrigation or sprinkler irrigation, and the concentration of the fertilizer solution is 0.5%-1%.

9. The method for standardized selection and breeding of authentic medicinal material seeds according to claim 1, characterized in that: In the pest control step, physical control uses insect-proof nets and yellow boards to lure and kill pests, and biological control releases natural enemies of pests. When the pests are serious, high-efficiency, low-toxicity, and low-residue pesticides are used for control.

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