Artificial propagation method of saccharum arundinaceum

Abstract

The invention relates to the technical field of plant propagation, in particular to an artificial propagation method of saccharum arundinaceum. The breeding method comprises the following steps: S1, preparing a culture medium; s2, preparing a seed treatment agent; s3, seed pretreatment; s4, sowing the seeds; s5, cultivating after sowing; s6, seedling culture; and S7, transplanting the seedlings. By means of the breeding method, the seed germination rate, the survival rate after transplanting and the seedling early-stage growth height of the saccharum arundinaceum are remarkably increased.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of plant propagation, in particular to an artificial propagation method of Eremochloa ophiuroides. BACKGROUND

[0002] Eremochloa ophiuroides, also known as Pishan Eremochloa, is a perennial tall herbaceous plant of the genus Saccharum in the family Poaceae. Its root system is well developed, with a depth of up to 1.5 meters, and its rhizome is short and can form large clusters. The plant stalk is usually 2 to 3 meters high, and the highest can reach 4 meters, with significant ecological and economic value. This species is naturally distributed in the mountainous areas of Xinjiang, China, and in the fixed dunes, gobi and sandy soil environments from northwest India, Central Asia to the Mediterranean region, with an altitude range of 1200 to 3000 meters. As a typical sand plant, Eremochloa ophiuroides has a large cluster of plants and well-developed basal leaves, with strong sand-blocking ability, and is considered an excellent sand-fixing plant, playing an important role in preventing land desertification and restoring the ecological system of sandy areas. Eremochloa ophiuroides mainly reproduces through seeds and rhizomes under natural conditions. Eremochloa ophiuroides has a unique geographical distribution and strong adaptability, with strong drought resistance, cold resistance and barren tolerance, but it is not resistant to heat and humidity, and there is no wild Eremochloa ophiuroides in low-altitude areas, and it is also difficult to survive in introduced test planting. SUMMARY

[0003] In view of the deficiencies of the prior art, the present application provides an artificial propagation method of Eremochloa ophiuroides.

[0004] The present application is realized by the following technical solutions: An artificial propagation method of Eremochloa ophiuroides, comprising the following steps: S1. Preparation of culture medium: S11. Dissolve polyvinyl alcohol in DMF, add chloroethyl isocyanate under nitrogen protection, stir for 15-20 min, add dibutyltin dilaurate, stir at 65-70℃ for 3-4 h, distill under reduced pressure, wash with ethanol, and dry under vacuum to obtain Cl-polyvinyl alcohol; S12. Mix the Cl-polyvinyl alcohol obtained in step S11 with DMSO, mix well, add eugenol and triethylamine under nitrogen protection, mix well, and react at 40-50℃ for 5-6 h, distill under reduced pressure, wash with ethanol, and dry under vacuum to obtain modified polyvinyl alcohol; S13. Dissolve hyaluronic acid in deionized water, add EDC·HCl and NHS, both at a concentration of 50 mmol / L, adjust the pH to 5.5 with 1 mol / L HCl, stir at room temperature in the dark for 30 min, add L-cysteine hydrochloride, adjust the pH to 4.75, and stir at room temperature in the dark for 5 h; S14. After the reaction in step S13 is completed, sequentially dialyze the product in a dark environment with an HCl solution having a pH of 5, an HCl solution containing 1% w / v NaCl having a pH of 5, and an HCl solution having a pH of 5 for 3 days, and freeze-dry the product to obtain SH-hyaluronic acid; S15. Add the modified polyvinyl alcohol obtained in step S12 to deionized water at a concentration of 0.2% by weight, heat to 60-70°C, and stir for 30 minutes to obtain liquid A. Dissolve the SH-hyaluronic acid obtained in step S14 in deionized water at a concentration of 0.5% by weight to obtain liquid B. Mix liquid A and liquid B, and add a photoinitiator LAP to a final concentration of 0.01% w / v. Irradiate with 365 nm ultraviolet light for 15-20 minutes. S16. After the reaction in step S15 is completed, wash the product in deionized water, store at 4°C overnight, and freeze-dry the product. Screen the product through a 20-mesh sieve, and mix the product with river sand, grass carbon, and vermiculite at a mass ratio of 2:5:3:2 to obtain a culture medium. S2. Preparation of a seed treatment agent: S21. Dry and grind the Fimbristylis littoralis, Raoulia australis, and Magnolia officinalis separately through an 80-100 mesh sieve. Mix the ground materials at a mass ratio of 3:2:1, and mix the mixture with 15-20 times the weight volume of deionized water. Sterilize the mixture at 121°C for 30 minutes, and cool the mixture to room temperature. Inoculate the mixture with 4 x 10 7 CFU / mL of activated Bacillus subtilis and Lactobacillus casei; S22. After inoculation in step S21, incubate the mixture at 37°C on a 150-200 rpm shaker for 48 hours. Centrifuge the mixture at 10,000 rpm for 15-20 minutes, pass the supernatant through a 0.22 μm filter, and mix the supernatant with gibberellin (GA3) at a ratio of 200 mg / L to obtain a seed treatment agent. S3. Seed pretreatment: Select full, undamaged, and non-shriveled seeds of the sand cattail, and immerse the seeds in the seed treatment agent after sterilization for 24 hours. Rinse the seeds with clean water. S4. Seed sowing: Fill the culture medium into a seedling pot (8 x 8 cm) after sterilization, and sow the seeds treated in step S3 in the center of the seedling pot, 2-3 seeds per pot. Cover the seeds with 0.5-1.0 cm of the culture medium, press the medium gently, and pour water through the medium. S5. Post-sowing cultivation: Place the seedling pots in an environment having a day temperature of 25-30°C and a night temperature of 15-20°C, and maintain the relative humidity of the air at 70-80% initially, and at 60% after germination. Keep the culture medium moist. S6. Seedling culture: After the seedlings grow 2-3 leaves, spray the seedlings with a 1000-fold solution of 50% w / w carbendazim wettable powder and a 1000-fold solution of 50% w / w methylthiophanate wettable powder alternately every 7 days. S7. Transplanting seedlings: when the seedlings have 4-5 leaves, gradually reduce watering, increase ventilation and direct light exposure time for 7-10 days, transplant in late spring, take the complete matrix soil lump when the seedlings are up, and plant in the pre-prepared sandy land or sandy soil according to the plant row spacing of 50 cm*50 cm, pour enough water for root fixation after planting, and carry out routine field management after planting.

[0005] Further, in step S11, the mass concentration of the polyvinyl alcohol in DMF is 15-20 mg / mL.

[0006] Further, in step S11, the mass ratio of the polyvinyl alcohol to chloroethyl isocyanate is 1.5-2:1.

[0007] Further, in step S11, the amount of dibutyltin dilaurate is 0.2-0.3wt% of chloroethyl isocyanate.

[0008] Further, in step S12, the mass concentration of the Cl-polyvinyl alcohol in DMSO is 20 mg / mL.

[0009] Further, in step S12, the mass ratio of the Cl-polyvinyl alcohol, eugenol and triethylamine is 1.6-1.8:1:0.7.

[0010] Further, in step S13, the mass concentration of the hyaluronic acid in deionized water is 4 mg / mL.

[0011] Further, in step S13, the mass ratio of the hyaluronic acid to L-cysteine hydrochloride is 1:2.

[0012] Further, in step S15, the mass ratio of the modified polyvinyl alcohol to SH-hyaluronic acid is 1:2-2.5.

[0013] Compared with the prior art, the present application has the following beneficial effects: The present application provides an artificial propagation method of psammophyte, which significantly improves the seed germination rate, survival rate after transplanting and seedling growth height of psammophyte, effectively overcomes the problems of difficult seed germination and fragile seedlings of psammophyte through the synergistic effect of specific cultivation substrate and seed treatment agent.

[0014] The core of the invention is to cultivate the substrate, which is a functional hydrogel material with interpenetrating network structure synthesized by chemical modification. First, the hydroxyl group of polyvinyl alcohol is reacted with the isocyanate group of chloroethyl isocyanate to introduce active chlorine atoms to obtain Cl-polyvinyl alcohol. Then, the substitution reaction of Cl-polyvinyl alcohol and the phenolic hydroxyl group of eugenol occurs under alkaline conditions to introduce the double bond structure of eugenol into the PVA molecular chain, providing a site for the subsequent "ene-thiol" click chemistry reaction. The present invention uses EDC catalysis to graft the thiol group of L-cysteine to hyaluronic acid to obtain thiolated hyaluronic acid. Hyaluronic acid itself has excellent water retention and biocompatibility. The modified polyvinyl alcohol is mixed with the SH-hyaluronic acid solution in the presence of a photoinitiator, and the two are crosslinked by efficient "ene-thiol" click chemistry under ultraviolet light irradiation to form a stable double network hydrogel with a three-dimensional porous structure, which can efficiently absorb water and dissolved nutrients and slowly release them in sandy substrates, providing a continuous, stable, and humid microenvironment for seed germination and seedling root growth. The addition of hydrogel particles can effectively improve the agglomeration of inorganic substrates such as river sand, increase the porosity, and promote root respiration and extension. In addition, the introduced eugenol has natural antibacterial and growth-promoting effects, which can help inhibit soil-borne pathogens and promote root health. Hyaluronic acid can stimulate plant root development and enhance absorption capacity. The seed treatment agent of the present invention uses a specific ratio of traditional Chinese medicine fermentation broth combined with gibberellin, selects Prunella vulgaris, Echinacea purpurea, and Magnolia officinalis for combination, and comprehensively utilizes the active ingredients of each medicinal material. Bacillus subtilis and Lactobacillus casei are used for compound fermentation. The microbial fermentation process can decompose and transform macromolecular polysaccharides and proteins in medicinal materials into small molecular active substances that are more easily absorbed, and also produces a large amount of microbial secondary metabolites, improving the biological activity and availability of traditional Chinese medicine ingredients. Gibberellin is a plant hormone that directly promotes seed germination and stem elongation, and has a synergistic effect with various active ingredients in the fermentation broth. The various growth regulators produced by fermentation and exogenous GA3 can more effectively activate the internal metabolism of the seed and break the dormancy of the sand cane seed. The small molecule nutrients in the fermentation broth can provide early energy for seed germination and improve the resistance to environmental stress. At the same time, the fermentation product components have antibacterial activity and can reduce disease infection. BRIEF DESCRIPTION OF DRAWINGS

[0015] In order to more clearly illustrate the technical solutions of the present application or prior art, the following will briefly introduce the drawings needed to be used in the embodiments or prior art description. Obviously, the drawings described below are only a part of the present application, and other drawings can also be obtained by those skilled in the art without creative labor.

[0016] Figure 1 Germination rate of the artificial propagation method described in Examples 1-3 and Comparative Examples 1-6; Figure 2 The seedling height of the artificial propagation method according to the embodiments 1-3 and the comparative examples 1-6 of the present application; Figure 3 The survival rate of the artificial propagation method according to the embodiments 1-3 and the comparative examples 1-6 of the present application. DETAILED DESCRIPTION

[0017] In order to make the objectives, technical solutions and advantages of the present application clearer, the present application will be further described in detail below with specific embodiments, but the present application is not limited to the following embodiments. It should be noted that, unless otherwise specified, the chemical reagents involved in the present application are purchased through commercial channels.

[0018] Embodiment 1: An artificial propagation method of a desert grass, comprising the following steps: S1. Preparation of the culture medium: S11. Dissolve 10 g of polyvinyl alcohol in 500 mL of DMF, add 5 g of chloroethyl isocyanate under nitrogen protection, stir for 20 min, add 15 mg of dibutyltin dilaurate, stir at 70°C for 4 h, distill under reduced pressure, wash with ethanol, and dry in vacuum to obtain Cl-polyvinyl alcohol; S12. Mix 10.8 g of the Cl-polyvinyl alcohol obtained in step S11 in 540 mL of DMSO, mix 6 g of eugenol and 4.2 g of triethylamine under nitrogen protection, react at 50°C for 6 h, distill under reduced pressure, wash with ethanol, and dry in vacuum to obtain modified polyvinyl alcohol; S13. Dissolve 2 g of hyaluronic acid in 500 mL of deionized water, add EDC·HCl and NHS, both at a concentration of 50 mmol / L, adjust the pH to 5.5 with 1 mol / L HCl, stir at room temperature in the dark for 30 min, add 4 g of L-cysteine hydrochloride, adjust the pH to 4.75, and stir at room temperature in the dark for 5 h; S14. After the reaction in step S13 is completed, sequentially dialyze in the dark for 3 days with HCl solution at pH=5, HCl solution containing 1% w / v NaCl at pH=5, and HCl solution at pH=5, and freeze-dry to obtain SH-hyaluronic acid; S15. Add 1 g of the modified polyvinyl alcohol obtained in step S12 to 500 mL of deionized water at a proportion of 0.2 wt%, heat to 70°C and stir for 30 min to obtain liquid A, add 2.5 g of the SH-hyaluronic acid obtained in step S14 to 500 mL of deionized water at a proportion of 0.5 wt% and stir to dissolve to obtain liquid B, mix liquid A and liquid B, add a photoinitiator LAP to a final concentration of 0.01% w / v, and irradiate with 365 nm ultraviolet light for 20 min; S16. After the reaction in step S15 is completed, soak and wash with deionized water, incubate overnight at 4°C, freeze dry, pass through a 20-mesh sieve, and mix evenly with river sand, peat moss and vermiculite in a mass ratio of 2:5:3:2 to obtain the cultivation substrate; S2. Preparation of seed treatment agent: S21. Wash and dry Prunella vulgaris, Echinacea purpurea, and Magnolia officinalis, then pulverize them separately and pass them through a 100-mesh sieve. Mix them in a mass ratio of 3:2:1, add 20 times their weight volume of deionized water, mix well, sterilize at 121℃ for 30 min, cool to room temperature, and then apply 4×10⁻⁶ ppm of each mixture. 7 Inoculate activated Bacillus subtilis and Lactobacillus casei at an inoculation rate of CFU / mL; S22. After inoculation in step S21, culture at 37℃ and 200 rpm in a shaker for 48 h, centrifuge at 10000 rpm for 20 min, filter the supernatant through a 0.22 μm filter membrane, add gibberellin (GA3) at a ratio of 200 mg / L and mix well to obtain the seed treatment agent; S3. Seed pretreatment: Select plump, undamaged, and shriveled sand sugarcane grass seeds. After disinfection, soak the seeds in the seed treatment agent for 24 hours, then rinse them with clean water. S4. Seed sowing: After sterilizing the culture substrate, fill the seedling pot (8×8cm), sow the S3 treated seeds in the center of the seedling pot, 3 seeds per pot, cover with substrate 1.0 cm, press lightly, and water thoroughly; S5. Post-sowing cultivation: Place the seedling pots in an environment that maintains a daytime temperature of 30℃ and a nighttime temperature of 20℃. Initially, maintain the relative humidity at 80%, and reduce it to 60% after germination. Keep the substrate moist but not waterlogged. S6. Seedling cultivation: After the seedlings have grown 3 leaves, spray them alternately every 7 days with 1000 times dilution of 50wt% carbendazim wettable powder and 1000 times dilution of 50wt% thiophanate-methyl wettable powder. S7. Transplanting of seedlings: When the seedlings have 5 leaves, gradually reduce watering and increase ventilation and direct sunlight exposure for 10 days. Transplant in late spring, keeping the seedlings with an intact substrate soil ball. Plant them in pre-prepared sandy soil or sandy loam at a spacing of 50 cm × 50 cm. Water thoroughly after planting and carry out routine field management after transplanting.

[0019] Example 2: An artificial propagation method for *Imperata cylindrica*, comprising the following steps: S1. Preparation of culture substrate: S11. Dissolve polyvinyl alcohol 9 g in DMF 600 mL, under nitrogen protection, add chloroethyl isocyanate 6 g, stir for 15 min, add dibutyltin dilaurate 12 mg, stir at 65℃ for 3 h, distill under reduced pressure, wash with ethanol, and dry in vacuum to obtain Cl-polyvinyl alcohol; S12. Mix the Cl-polyvinyl alcohol 8 g obtained in step S11 in DMSO 400 mL, under nitrogen protection, mix in eugenol 5 g and triethylamine 3.5 g, react at 40℃ for 5 h, distill under reduced pressure, wash with ethanol, and dry in vacuum to obtain modified polyvinyl alcohol; S13. Take hyaluronic acid 2 g, dissolve in deionized water 500 mL, add EDC·HCl and NHS, both at a concentration of 50 mmol / L, adjust the pH to 5.5 with 1 mol / L HCl, stir at room temperature in the dark for 30 min, add L-cysteine hydrochloride 4 g, adjust the pH to 4.75, and stir at room temperature in the dark for 5 h; S14. After the reaction in step S13 is completed, sequentially dialyze in HCl solution at pH=5, HCl solution containing 1% w / v NaCl at pH=5, and HCl solution at pH=5 in the dark for 3 days, and freeze-dry to obtain SH-hyaluronic acid; S15. Add the modified polyvinyl alcohol 1 g obtained in step S12 to deionized water 500 mL at a rate of 0.2 wt%, heat to 60℃ and stir for 30 min to obtain liquid A, add the SH-hyaluronic acid 2 g obtained in step S14 to deionized water 400 mL at a rate of 0.5 wt% and stir to dissolve to obtain liquid B, mix liquids A and B, add a photoinitiator LAP to a final concentration of 0.01% w / v, and irradiate with 365 nm ultraviolet light for 15 min; S16. After the reaction in step S15 is completed, soak and wash in deionized water, and store at 4℃ overnight, then freeze-dry and pass through a 20 mesh sieve, mix with river sand, grass charcoal and vermiculite at a mass ratio of 2:5:3:2 to obtain a cultivation substrate; S2. Preparation of seed treatment agent: S21. Wash and dry the Spica Prunellae, Echinacea purpurea and Magnolia officinalis, crush and pass through an 80 mesh sieve, mix at a mass ratio of 3:2:1, mix in 15 times the weight volume of deionized water, sterilize at 121℃ for 30 min, cool to room temperature, and inoculate with activated Bacillus subtilis and Lactobacillus casei at an inoculum of 4×10 7 CFU / mL; S22. After inoculation in step S21, cultivate at 37℃ on a 150 rpm shaker for 48 h, centrifuge at 10000 rpm for 15 min, pass the supernatant through a 0.22 μm filter membrane, and mix in gibberellin (GA3) at a rate of 200 mg / L to obtain a seed treatment agent; S3. Seed pretreatment: Select full, non-injured, non-shriveled sand cattail seeds, soak the seeds in the seed treatment agent for 24 h after disinfection, and then rinse them clean with clean water; S4. Seed sowing: After sterilizing and disinfecting the cultivation substrate, fill it into the seedling pots (8x8 cm), sow the seeds treated in S3 in the center of the seedling pots, 2 seeds per pot, cover the substrate with 0.5 cm, lightly press, and pour water thoroughly; S5. Post-seeding cultivation: Place the seedling pots in an environment with a day temperature of 25°C and a night temperature of 15°C, and maintain the relative humidity of the air at 70% initially, and reduce it to 60% after germination, and keep the substrate moist; S6. Seedling culture: After the seedlings grow 2 leaves, spray them with 50wt% carbendazim wettable powder 1000 times liquid and 50wt% methylthiophanate wettable powder 1000 times liquid alternately every 7 days; S7. Seedling transplanting: When the seedlings have 4 leaves, gradually reduce watering, increase ventilation, and increase direct light exposure time for 7 days, transplant in late spring, take the entire substrate soil clumps when transplanting, and plant them in the prepared sandy soil or sandy loam soil with a plant spacing of 50 cm x 50 cm, pour enough water to fix the roots after planting, and perform routine field management after planting.

[0020] Example 3: A method for artificially propagating sand cattail, comprising the following steps: S1. Preparation of cultivation substrate: S11. Dissolve 9 g of polyvinyl alcohol in 500 mL of DMF, add 5 g of chloroethyl isocyanate under nitrogen protection, stir for 15 min, add 12 mg of dibutyltin dilaurate, stir at 68°C for 3 h, distill under reduced pressure, wash with ethanol, and vacuum dry to obtain Cl-polyvinyl alcohol; S12. Mix 8 g of Cl-polyvinyl alcohol obtained in step S11 in 400 mL of DMSO, mix 4.5 g of eugenol and 3.15 g of triethylamine, and react at 40°C for 5 h under nitrogen protection, distill under reduced pressure, wash with ethanol, and vacuum dry to obtain modified polyvinyl alcohol; S13. Dissolve 2 g of hyaluronic acid in 500 mL of deionized water, add EDC·HCl and NHS, both at a concentration of 50 mmol / L, adjust the pH to 5.5 with 1 mol / L HCl, stir at room temperature for 30 min in the dark, add 4 g of L-cysteine hydrochloride, adjust the pH to 4.75, and stir at room temperature for 5 h in the dark; S14. After the reaction in step S13 is completed, sequentially dialyze with pH=5 HCl solution, 1% w / v NaCl pH=5 HCl solution, and pH=5 HCl solution in the dark for 3 days, and freeze-dry to obtain SH-hyaluronic acid; S15. The modified polyvinyl alcohol 1 g obtained in step S12 was added to 500 mL of deionized water at 0.2 wt%, heated to 60-70°C and stirred for 30 min to obtain liquid A. The SH-hyaluronic acid 2.2 g obtained in step S14 was added to 440 mL of deionized water at 0.5 wt% and stirred to dissolve to obtain liquid B. Liquid A and liquid B were mixed, and a photoinitiator LAP was added to a final concentration of 0.01% w / v, and irradiated with 365 nm ultraviolet light for 18 min; S16. After the reaction in step S15 was completed, the product was soaked and washed with deionized water, stored at 4°C overnight, freeze-dried, and sieved through a 20 mesh screen. The product was mixed with river sand, grass charcoal and vermiculite at a mass ratio of 2:5:3:2 to obtain a cultivation medium; S2. Preparation of seed treatment agent: S21. The Fructus Pruni, Echinacea purpurea and Magnolia officinalis were washed, dried and ground through a 90 mesh screen. The three were mixed at a mass ratio of 3:2:1, added to 18 times the weight volume of deionized water, sterilized at 121°C for 30 min, and cooled to room temperature. Each was inoculated with activated Bacillus subtilis and Lactobacillus casei at an inoculum of 4x10 7 CFU / mL; S22. After inoculation in step S21, the mixture was incubated at 37°C with shaking at 180 rpm for 48 h, centrifuged at 10,000 rpm for 18 min, and the supernatant was filtered through a 0.22 μm filter membrane. Gibberellin (GA3) was added at a ratio of 200 mg / L to obtain a seed treatment agent; S3. Seed pretreatment: Full, undamaged, non-shriveled seeds of Puccinellia tanakae were selected, sterilized and soaked in the seed treatment agent for 24 h, then rinsed with clean water; S4. Seed sowing: The cultivation medium was sterilized and filled into seedling pots (8x8 cm), and the seeds treated in S3 were sown in the center of the pots, 3 seeds per pot, covered with 0.8 cm of medium, lightly pressed, and watered thoroughly; S5. Post-sowing cultivation: The seedling pots were placed in an environment with a day temperature of 28°C and a night temperature of 18°C, with an initial relative humidity of 75%, which was reduced to 60% after germination, and the medium was kept moist; S6. Seedling culture: After the seedlings grew 3 leaves, they were sprayed every 7 days with a 1000-fold solution of 50 wt% carbendazim wettable powder and a 1000-fold solution of 50 wt% methylthiophanate wettable powder alternately; S7. Seedling transplanting: When the seedlings had 5 leaves, the watering was gradually reduced, the ventilation and direct light exposure time were increased, and this was continued for 8 days. The seedlings were transplanted in late spring, with the entire medium soil clumps, and were planted in prepared sandy soil or sandy loam soil at a plant spacing of 50 cm x 50 cm. After planting, the roots were watered thoroughly, and routine field management was performed.

[0021] Comparative Example 1 differs from Example 1 only in that polyvinyl alcohol is used instead of modified polyvinyl alcohol.

[0022] Comparative Example 2 differs from Example 1 only in that hyaluronic acid is used instead of SH-hyaluronic acid.

[0023] Comparative Example 3 differs from Example 1 only in that Echinacea and Magnolia are not added, and only Spica Prunellae is used.

[0024] Comparative Example 4 differs from Example 1 only in that Spica Prunellae, Echinacea and Magnolia are washed, dried, ground through an 80-100 mesh sieve, mixed with 15-20 times the weight volume of deionized water, soaked for 1 h, decocted, boiled for 30 min, filtered, and the above operation repeated, the two times of decocting liquid combined, filtered through a 0.22 μm filter membrane, and gibberellin (GA3) added at a ratio of 200 mg / L to obtain a seed treatment agent.

[0025] Comparative Example 5 differs from Example 1 only in that Bacillus subtilis is used alone.

[0026] Comparative Example 6 differs from Example 1 only in that gibberellin (GA3) is added to water at a ratio of 200 mg / L to obtain a seed treatment agent.

[0027] Experimental Example 1: According to the methods described in Examples 1-3 and Comparative Examples 1-6, the germination rate of seedlings of Sporobolus virginicus was counted after 7 days of sowing, and the results are shown in Table 1. Figure 1

[0028] Figure 1 The results show that the germination rate of Examples 1-3 is significantly higher than that of Comparative Examples 1-6. Comparative Example 1 uses polyvinyl alcohol instead of modified polyvinyl alcohol, which is not modified and lacks eugenol groups. Comparative Example 2 uses hyaluronic acid instead of SH-hyaluronic acid, which cannot form effective cross-linking and reduces the effect of promoting seed germination. Comparative Example 3 does not add Echinacea and Magnolia, and only uses Spica Prunellae. Comparative Example 4 extracts the traditional Chinese medicine by decocting, but does not ferment the traditional Chinese medicine. Comparative Example 5 uses only Bacillus subtilis. Comparative Example 6 only uses gibberellin to treat the seeds. The active ingredients of the seed treatment agents of Comparative Examples 3-6 are reduced, the availability is reduced, and the effect of promoting seed germination is reduced.

[0029] Experimental Example 2: According to the methods described in Examples 1-3 and Comparative Examples 1-6, the seedling height of Sporobolus virginicus was counted after 15 days of sowing, and the results are shown in Table 2. Figure 2

[0030] Figure 2 ​​The results showed that the seedling height of *Imperata cylindrica* in Examples 1-3 was significantly higher than that in Comparative Examples 1-6. Comparative Example 1 used polyvinyl alcohol instead of modified polyvinyl alcohol, but did not modify the polyvinyl alcohol and lacked eugenol groups. Comparative Example 2 used hyaluronic acid instead of SH-hyaluronic acid, which could not form effective cross-links and reduced the effect of promoting plant growth. Comparative Example 3 did not add *Echinacea purpurea* and *Magnolia officinalis*, but only used *Prunella vulgaris*. Comparative Example 4 used the Chinese herbal medicine by soaking and boiling without fermenting it. Comparative Example 5 only used *Bacillus subtilis*. Comparative Example 6 only used gibberellin to treat the seeds. The active ingredients of the seed treatment agents in Comparative Examples 3-6 decreased, the utilization rate decreased, and the effect of promoting plant growth decreased.

[0031] Experimental Example 3: Following the methods described in Examples 1-3 and Comparative Examples 1-6, the survival rate of *Imperata cylindrica* seedlings was calculated 30 days after transplanting. The results are as follows: Figure 3 As shown.

[0032] Figure 3 The results showed that the survival rate of *Imperata cylindrica* in Examples 1-3 was significantly higher than that in Comparative Examples 1-6. Comparative Example 1 used polyvinyl alcohol instead of modified polyvinyl alcohol, but did not modify the polyvinyl alcohol and lacked eugenol groups. Comparative Example 2 used hyaluronic acid instead of SH-hyaluronic acid, which could not form effective cross-links and reduced the effect of promoting plant survival. Comparative Example 3 did not add *Echinacea purpurea* and *Magnolia officinalis*, but only used *Prunella vulgaris*. Comparative Example 4 used the Chinese herbal medicine by soaking and boiling without fermentation. Comparative Example 5 only used *Bacillus subtilis*. Comparative Example 6 only used gibberellin to treat seeds. The active ingredients of the seed treatment agents in Comparative Examples 3-6 decreased, the utilization rate decreased, and the effect of promoting plant survival decreased.

[0033] Those skilled in the art should understand that the discussion of any of the above embodiments is merely exemplary and is not intended to imply that the scope of the invention is limited to these examples; within the framework of the invention, the technical features of the above embodiments or different embodiments can also be combined, the steps can be implemented in any order, and there are many other variations of the different aspects of the invention as described above, which are not provided in detail for the sake of brevity.

Claims

1. A method for the artificial propagation of *Imperata cylindrica*, characterized in that, Includes the following steps: S1. Preparation of culture substrate: S11. Polyvinyl alcohol is dissolved in DMF, chloroethyl isocyanate is added, stirred, dibutyltin dilaurate is added, stirred to react, distilled under reduced pressure, washed, and dried to obtain Cl-polyvinyl alcohol; S12. Add the Cl-polyvinyl alcohol obtained in step S11 to DMSO and mix well. Add eugenol and triethylamine and mix well. React, distill under reduced pressure, wash, and dry to obtain modified polyvinyl alcohol. S13. Dissolve hyaluronic acid in deionized water, add EDC·HCl and NHS, adjust the pH, stir, add L-cysteine ​​hydrochloride, adjust the pH, and stir. S14. After the reaction in step S13 is completed, dialyze and freeze-dry to obtain SH-hyaluronic acid; S15. Add the modified polyvinyl alcohol obtained in step S12 to deionized water to obtain solution A. Add the SH-hyaluronic acid obtained in step S14 to deionized water to obtain solution B. Mix solution A and solution B, add photoinitiator LAP, and irradiate with ultraviolet light. S16. After the reaction in step S15 is completed, wash, freeze-dry, pass through a sieve, and mix evenly with river sand, peat moss and vermiculite in a mass ratio of 2:5:3:2 to obtain the cultivation substrate; S2. Preparation of seed treatment agent: S21. Wash and dry the Prunella vulgaris, Echinacea purpurea and Magnolia officinalis, pulverize them and pass them through a sieve, mix them in a mass ratio of 3:2:1, add them to deionized water, sterilize, cool, and inoculate with Bacillus subtilis and Lactobacillus casei. S22. After inoculation in step S21, culture, centrifuge, filter, add gibberellin and mix well to obtain seed treatment agent; S3. Seed pretreatment: Select sand-grown sugarcane grass seeds, disinfect the seeds, soak them in seed treatment agent, and rinse with clean water; S4. Seed sowing: Fill the seedling pot with the culture substrate, sow the S3 treated seeds in the center of the seedling pot, cover with substrate, and water thoroughly; S5. Post-sowing cultivation; S6. Seedling cultivation; S7. Transplanting of seedlings.

2. The method for artificial propagation of *Imperata cylindrica* according to claim 1, characterized in that, In step S11, the mass ratio of polyvinyl alcohol to chloroethyl isocyanate is 1.5-2:

1.

3. The method for artificial propagation of *Imperata cylindrica* according to claim 2, characterized in that, In step S11, the amount of dibutyltin dilaurate used is 0.2-0.3 wt% of chloroethyl isocyanate.

4. The method for artificial propagation of *Imperata cylindrica* according to claim 3, characterized in that, In step S12, the mass ratio of Cl-polyvinyl alcohol, eugenol and triethylamine is 1.6-1.8:1:0.

7.

5. The method for artificial propagation of *Imperata cylindrica* according to claim 4, characterized in that, In step S13, the mass ratio of hyaluronic acid to L-cysteine ​​hydrochloride is 1:

2.

6. The method for artificial propagation of *Imperata cylindrica* according to claim 5, characterized in that, In step S15, the mass ratio of the modified polyvinyl alcohol to SH-hyaluronic acid is 1:2-2.5.

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