Method for freshwater cultivation of mangrove plant bruguiera gymnorrhiza
By using a seed soaking solution of Veratrum nigrum leaf extract and a compound fertilizer containing Bacillus curvatureii, the problems of ion balance and trace element deficiency in female sea olives in freshwater environments were solved, resulting in high germination and high survival rates for female sea olive seedlings.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- HAINAN ACAD OF FORESTRY SCI (HAINAN ACAD OF MANGROVE RES)
- Filing Date
- 2024-12-31
- Publication Date
- 2026-05-15
AI Technical Summary
In existing technologies, the cultivation of female sea olives in freshwater environments faces problems such as ion imbalance, trace element deficiency, and insufficient microbial community, resulting in low germination rate, poor growth, and difficulty in normal growth in freshwater environments.
The seed soaking solution and compound fertilizer are used for treatment. The seed soaking solution is formulated with extracts from Veratrum nigrum leaves, and the compound fertilizer contains poultry and livestock manure, Livistona chinensis leaves and Bacillus curvatureis bacterial solution to establish a stable ion balance and microbial community, which promotes seed germination and seedling growth.
It improved the germination rate and survival rate of female sea olives in freshwater environments, with a germination rate of over 97%, and seedlings reaching a height of 57cm after 6 months of cultivation, with a survival rate of over 96%.
Smart Images
Figure CN119769346B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of mangrove seedling technology, specifically to a freshwater cultivation method for the mangrove plant *Gynostemma pentaphyllum*. Background Technology
[0002] Mangrove plants typically grow in the intertidal zone along the coast, having long adapted to marine or brackish water environments characterized by high salinity, tidal variations, and specific soil conditions. Current technologies for mangrove cultivation primarily focus on their native marine or brackish water environments. However, with environmental changes and the increasing demand for diversified plant resources, exploring methods for cultivating mangrove plants in freshwater environments is of significant importance. The adaptability of mangrove plants to freshwater cultivation varies by species. For example, studies have shown that certain mangrove species, such as *Aegiceras corniculatum*, *Sonneratia apetala*, and *Sonneratia caseolaris*, can grow normally in freshwater environments, exhibiting good adaptability and growth performance. Other species, such as *Kandelia candel* and *Excoecaria agallocha*, have lower survival rates and poorer growth under freshwater conditions.
[0003] *Avicennia marina*, an important mangrove plant, thrives in warm, humid climates and is found primarily in coastal and salt marshland areas, ranging from high to low tide. As a pioneer species of mangroves, *Avicennia marina* plays a crucial role in wind and wave protection, siltation and beach conservation, embankment stabilization, and ecological balance regulation. Currently, cultivation of *Avicennia marina* is mainly concentrated in brackish water environments. Due to its physiological characteristics and ecological requirements, traditional cultivation methods face numerous challenges in freshwater environments. For example, the significant difference in osmotic pressure between freshwater and seawater leads to an imbalance in water absorption and loss in plant cells, affecting normal physiological functions. Furthermore, *Avicennia marina* has adapted to high-salinity environments through long-term evolution, and its ion balance mechanisms and metabolic pathways are not adapted to freshwater environments, thus affecting key physiological processes such as nutrient absorption and photosynthesis. Moreover, compared to seawater, freshwater environments lack certain specific trace elements or microbial communities necessary for the growth of *Avicennia marina*, further increasing the difficulty of cultivation. Summary of the Invention
[0004] In view of the shortcomings of the existing technology, in order to overcome the difficulties in freshwater cultivation of the mangrove plant *Rhizophora spp.*, the inventors conducted in-depth research and innovation and proposed a freshwater cultivation method for the mangrove plant *Rhizophora spp.*.
[0005] The technical solution of this invention is as follows:
[0006] A freshwater cultivation method for the mangrove plant *Gynostemma pentaphyllum* includes the following steps:
[0007] (1) Seed treatment
[0008] Take female seeds of sea olives and soak them in a seed soaking solution; after soaking, sterilize them and air dry them in the shade.
[0009] The soaking solution is a 20-30% (w / w) extract of Veratrum nigrum leaf (prepared with water);
[0010] The preparation method of the veratrum leaf extract includes: taking fresh veratrum leaves, drying and pulverizing them, extracting them with petroleum ether at 60-70°C with stirring, filtering, extracting the filter residue with ethyl acetate at 70-75°C, collecting the ethyl acetate extract, and recovering the ethyl acetate under reduced pressure to obtain the veratrum leaf extract.
[0011] (2) Sowing
[0012] Prepare a seedling substrate and sow the seeds in a seedling bag containing the substrate; the seedling substrate contains sandy loam soil, poultry and livestock manure, fermented products, and phosphate fertilizer;
[0013] The fermentation formula includes: poultry and livestock manure, palm leaves, urea, and Bacillus curvatureii bacterial solution;
[0014] The method for making fermented products includes taking poultry and livestock manure, palm leaves, urea, and Bacillus flexuralis bacterial solution, spreading them out, sprinkling water and mixing them evenly, piling them into long strips, and fermenting for 3-5 days;
[0015] (3) Seedling management
[0016] Water regularly with fresh water and fertilize.
[0017] Preferably, the mass ratio of sandy loam soil to poultry and livestock manure, fermentation products, and phosphate fertilizer is 70-80:2-5:10-17:8-10.
[0018] Preferably, the fermentation formula, by weight, includes: 75-80 parts of poultry and livestock manure, 16.5-21 parts of palm leaves, 1-0.5 parts of urea, and 2-3 parts of Bacillus curvatureii bacterial solution.
[0019] Preferably, the soaking time is 3-5 days.
[0020] Preferably, the preparation method of the veratrum leaf extract includes: taking fresh veratrum leaves, drying and pulverizing them, extracting them with petroleum ether at 60-70°C with stirring for 5-7 hours, filtering, extracting the filter residue with ethyl acetate at 70-75°C for 8-10 hours, collecting the ethyl acetate extract, and recovering the ethyl acetate under reduced pressure to obtain the veratrum leaf extract.
[0021] Preferably, the poultry and livestock manure is dried cow manure.
[0022] Preferably, the soaking solution is a 30% (w / w) extract of Veratrum nigrum leaves.
[0023] Preferably, the mass ratio of sandy loam soil to poultry and livestock manure, fermentation products, and phosphate fertilizer is 80:2:10:10.
[0024] Preferably, the preparation method of Veratrum leaf extract is as follows: take fresh Veratrum leaves, dry and pulverize them, extract with petroleum ether at 60°C with stirring for 5 hours, filter, extract the filter residue with ethyl acetate at 70°C for 10 hours, collect the ethyl acetate extract, and recover the ethyl acetate under reduced pressure to obtain Veratrum leaf extract.
[0025] The preferred method for seedling management is as follows: water once a day with diluted water for the first two months of seedling cultivation, and then water once a week thereafter. Apply fertilizer once when the seedlings are two months old.
[0026] Compared with the prior art, the beneficial results of the present invention are as follows:
[0027] This invention introduces a specific seed soaking solution that helps female sea olives re-establish a relatively stable ion balance and metabolic pattern in a freshwater environment, promoting seed germination, ensuring nutrient absorption, and laying the foundation for subsequent seedling growth. Furthermore, to compensate for the lack of specific trace elements and microbial communities in freshwater environments, a compound fertilizer rich in nutrients and beneficial microorganisms has been specially developed. This fertilizer not only provides the trace elements needed for the growth of female sea olives but also improves the soil's ecological structure by introducing specific microbial communities and nutrient promoters, creating a more favorable microenvironment for the growth of female sea olives.
[0028] This invention effectively guarantees and promotes the germination rate, seedling height, and survival rate of female sea olives in freshwater environments. The germination rate is as high as 97% or more, the seedling height reaches 57cm after 6 months of cultivation, and the survival rate reaches 96% or more. Attached Figure Description
[0029] Figure 1 The seedlings cultivated in this embodiment of the invention. After removing the seedling container, the root ball is surrounded by a well-developed root system in a net-like pattern. Detailed Implementation
[0030] To enable those skilled in the art to better understand the technical content of the present invention, the present invention will be further described below in conjunction with specific embodiments and accompanying drawings.
[0031] Unless otherwise specified, water in the following examples refers to fresh water.
[0032] Example 1: A freshwater cultivation method for the mangrove plant *Gynostemma pentaphyllum*.
[0033] (1) Seed treatment
[0034] Harvest mature female fruits of the sea olive (the pericarp changes from grayish-green to yellowish-green). Soak the harvested fruits in water for about 3 hours until the pericarp detaches from the seed (also known as the hypocotyl). Remove the pericarp floating on the surface and rinse the seeds 2-3 times with clean water. Soak the seeds in a 30% (w / w) veratrum leaf extract for 5 days.
[0035] Preparation method of Veratrum leaf extract: Take fresh Veratrum leaves, dry and pulverize them, extract with petroleum ether at 60℃ for 5 hours with stirring, filter, extract the filter residue with ethyl acetate at 70℃ for 10 hours, collect the ethyl acetate extract, and recover the ethyl acetate under reduced pressure to obtain Veratrum leaf extract.
[0036] Disinfect the female seeds of sea olives that have undergone soaking treatment by soaking them in a 500-fold dilution of chlorothalonil solution for 3 to 8 minutes, then remove and air dry them before sowing.
[0037] (2) Sowing
[0038] Seedling substrate: a mixture of sandy loam soil, dried cow manure, fermented material, and phosphate fertilizer (superphosphate, available phosphorus P2O5 ≥ 12%, calcium ≥ 16%), in a mass ratio of 80:2:10:10. Sow seeds in seedling bags containing the substrate, and place the seedling bags on ordinary sandy loam soil. Two weeks after sowing, count the germination rate (with radicle length greater than 3 mm as the germination indicator, n = 30).
[0039] The fermentation formula is as follows (parts by weight): 75 parts dried cow dung, 21 parts fan palm leaves, 1 part urea, and 2 parts Bacillus flexus bacterial solution (ATCC 49095). The preparation method involves taking dried cow dung, fan palm leaves, urea, and Bacillus flexus bacterial solution (containing 10... 7 (cfu / mL), spread out, sprinkle with water and mix well (the amount of water should be such that when you squeeze the mixture in your hand, water drips out), pile it into a 1.0m×0.5m×0.2m long strip, ferment for 5 days, turning the pile once a day.
[0040] (3) Seedling management
[0041] For the first two months of seedling cultivation, water once a day (with fresh water), and then once a week thereafter. When the seedlings are two months old, apply fertilizer solution once (5% NPK balanced compound fertilizer).
[0042] Example 2: A freshwater cultivation method for the mangrove plant *Gynostemma pentaphyllum*.
[0043] The technical parameters in the preparation method of Veratrum nigrum leaf extract can be adjusted within a certain range. The following is one example.
[0044] (1) Seed treatment
[0045] Harvest mature female fruits of the sea olive (the pericarp changes from grayish-green to yellowish-green). Soak the harvested fruits in water for about 3 hours until the pericarp detaches from the seed (also known as the hypocotyl). Remove the pericarp floating on the surface and rinse the seeds 2-3 times with clean water. Soak the seeds in a 30% (w / w) veratrum leaf extract for 3 days.
[0046] Preparation method of Veratrum leaf extract: Take fresh Veratrum leaves, dry and pulverize them, extract with petroleum ether at 70℃ for 7 hours with stirring, filter, extract the filter residue with ethyl acetate at 75℃ for 8 hours, collect the ethyl acetate extract, and recover the ethyl acetate under reduced pressure to obtain Veratrum leaf extract.
[0047] Disinfect the female seeds of sea olives that have undergone soaking treatment by soaking them in a 500-fold dilution of chlorothalonil solution for 3 to 8 minutes, then remove and air dry them before sowing.
[0048] (2) Sowing
[0049] Prepare the seedling substrate: a mixture of sandy loam soil, dried cow manure, fermented material, and phosphate fertilizer (superphosphate, available phosphorus P2O5 ≥ 12%, calcium ≥ 16%), in a mass ratio of 80:2:10:10. Sow the seeds in seedling bags containing the substrate, and place the seedling bags on ordinary sandy loam soil. Two weeks after sowing, count the germination rate (with radicle length greater than 3 mm as the germination indicator, n = 30).
[0050] The fermentation formula is as follows (parts by weight): 75 parts dried cow dung, 21 parts fan palm leaves, 1 part urea, and 2 parts Bacillus flexus bacterial solution. The preparation method involves taking dried cow dung, fan palm leaves, urea, and Bacillus flexus bacterial solution (containing 10% bacteria). 7 (cfu / mL), spread out, sprinkle with water and mix well (the amount of water should be such that when you squeeze the mixture in your hand, water drips out), pile it into a 1.0m×0.5m×0.2m long strip, ferment for 3 days, turning the pile once a day.
[0051] (3) Seedling management
[0052] For the first two months of seedling cultivation, water once a day (with fresh water), and then once a week thereafter. When the seedlings are two months old, apply fertilizer solution once (5% NPK balanced compound fertilizer).
[0053] Example 3: A freshwater cultivation method for the mangrove plant *Gynostemma pentaphyllum*.
[0054] The technical parameters in the fermentation preparation method can be adjusted within a certain range. The following is one example.
[0055] (1) Seed treatment
[0056] Harvest mature female fruits of the sea olive (the pericarp changes from grayish-green to yellowish-green). Soak the harvested fruits in water for about 3 hours until the pericarp detaches from the seed (also known as the hypocotyl). Remove the pericarp floating on the surface and rinse the seeds 2-3 times with clean water. Soak the seeds in a 30% (w / w) veratrum leaf extract for 5 days.
[0057] Preparation method of Veratrum leaf extract: Take fresh Veratrum leaves, dry and pulverize them, extract with petroleum ether at 60℃ for 5 hours with stirring, filter, extract the filter residue with ethyl acetate at 70℃ for 10 hours, collect the ethyl acetate extract, and recover the ethyl acetate under reduced pressure to obtain Veratrum leaf extract.
[0058] Disinfect the female seeds of sea olives that have undergone soaking treatment by soaking them in a 500-fold dilution of chlorothalonil solution for 3 to 8 minutes, then remove and air dry them before sowing.
[0059] (2) Sowing
[0060] Seedling substrate: a mixture of sandy loam soil, dried cow manure, fermented material, and phosphate fertilizer (superphosphate, available phosphorus P2O5 ≥ 12%, calcium ≥ 16%), in a mass ratio of 80:2:10:10. Sow seeds in seedling bags containing the substrate, and place the seedling bags on ordinary sandy loam soil. Two weeks after sowing, count the germination rate (with radicle length greater than 3 mm as the germination indicator, n = 30).
[0061] The fermentation formula is as follows (parts by weight): 80 parts dried cow dung, 16.5 parts fan palm leaves, 0.5 parts urea, and 3 parts Bacillus flexus bacterial solution. The preparation method involves taking dried cow dung, fan palm leaves, urea, and Bacillus flexus bacterial solution (containing 10% bacteria). 7 (cfu / mL), spread out, sprinkle with water and mix well (the amount of water should be such that when you squeeze the mixture in your hand, water drips out), pile it into a 1.0m×0.5m×0.2m long strip, ferment for 5 days, turning the pile once a day.
[0062] (3) Seedling management
[0063] For the first two months of seedling cultivation, water once a day (with fresh water), and then once a week thereafter. When the seedlings are two months old, apply fertilizer solution once (5% NPK balanced compound fertilizer).
[0064] Example 4: A freshwater cultivation method for the mangrove plant *Gynostemma pentaphyllum*
[0065] The formula and ratio of the seedling substrate can be adjusted appropriately. The following is one example.
[0066] (1) Seed treatment
[0067] Harvest mature female fruits of the sea olive (the pericarp changes from grayish-green to yellowish-green). Soak the harvested fruits in water for about 3 hours until the pericarp detaches from the seed (also known as the hypocotyl). Remove the pericarp floating on the surface and rinse the seeds 2-3 times with clean water. Soak the seeds in a 30% (w / w) veratrum leaf extract for 5 days.
[0068] Preparation method of Veratrum leaf extract: Take fresh Veratrum leaves, dry and pulverize them, extract with petroleum ether at 60℃ for 5 hours with stirring, filter, extract the filter residue with ethyl acetate at 70℃ for 10 hours, collect the ethyl acetate extract, and recover the ethyl acetate under reduced pressure to obtain Veratrum leaf extract.
[0069] Disinfect the female seeds of sea olives that have undergone soaking treatment by soaking them in a 500-fold dilution of chlorothalonil solution for 3 to 8 minutes, then remove and air dry them before sowing.
[0070] (2) Sowing
[0071] Seedling substrate: a mixture of sandy loam soil, dried cow manure, fermented material, and phosphate fertilizer (superphosphate, available phosphorus P2O5 ≥ 12%, calcium ≥ 16%), in a mass ratio of 70:5:17:8. Sow seeds in seedling bags containing the substrate, and place the seedling bags on ordinary sandy loam soil. Two weeks after sowing, count the germination rate (with radicle length greater than 3 mm as the germination indicator, n = 30).
[0072] The fermentation formula is as follows (parts by weight): 75 parts dried cow dung, 21 parts fan palm leaves, 1 part urea, and 2 parts Bacillus flexus bacterial solution. The preparation method involves taking dried cow dung, fan palm leaves, urea, and Bacillus flexus bacterial solution (containing 10% bacteria). 7 (cfu / mL), spread out, sprinkle with water and mix well (the amount of water should be such that when you squeeze the mixture in your hand, water drips out), pile it into a 1.0m×0.5m×0.2m long strip, ferment for 5 days, turning the pile once a day.
[0073] (3) Seedling management
[0074] For the first two months of seedling cultivation, water once a day (with fresh water), and then once a week thereafter. When the seedlings are two months old, apply fertilizer solution once (5% NPK balanced compound fertilizer).
[0075] Example 5: A freshwater cultivation method for the mangrove plant *Gynostemma pentaphyllum*.
[0076] In this embodiment, the amount of Veratrum nigrum leaf extract used is 20%.
[0077] (1) Seed treatment
[0078] Harvest mature female fruits of the sea olive (the pericarp changes from grayish-green to yellowish-green). Soak the harvested fruits in water for about 3 hours until the pericarp detaches from the seed (also known as the hypocotyl). Remove the pericarp floating on the surface and rinse the seeds 2-3 times with clean water. Soak the seeds in a 20% (w / w) veratrum leaf extract for 5 days.
[0079] Preparation method of Veratrum leaf extract: Take fresh Veratrum leaves, dry and pulverize them, extract with petroleum ether at 60℃ for 5 hours with stirring, filter, extract the filter residue with ethyl acetate at 70℃ for 10 hours, collect the ethyl acetate extract, and recover the ethyl acetate under reduced pressure to obtain Veratrum leaf extract.
[0080] Disinfect the female seeds of sea olives that have undergone soaking treatment by soaking them in a 500-fold dilution of chlorothalonil solution for 3 to 8 minutes, then remove and air dry them before sowing.
[0081] (2) Sowing
[0082] Seedling substrate: a mixture of sandy loam soil, dried cow manure, fermented material, and phosphate fertilizer (superphosphate, available phosphorus P2O5 ≥ 12%, calcium ≥ 16%), in a mass ratio of 80:2:10:10. Sow seeds in seedling bags containing the substrate, and place the seedling bags on ordinary sandy loam soil. Two weeks after sowing, count the germination rate (with radicle length greater than 3 mm as the germination indicator, n = 30).
[0083] The fermentation formula is as follows (parts by weight): 75 parts dried cow dung, 21 parts fan palm leaves, 1 part urea, and 2 parts Bacillus flexus bacterial solution. The preparation method involves taking dried cow dung, fan palm leaves, urea, and Bacillus flexus bacterial solution (containing 10% bacteria). 7 (cfu / mL), spread out, sprinkle with water and mix well (the amount of water should be such that when you squeeze the mixture in your hand, water drips out), pile it into a 1.0m×0.5m×0.2m long strip, ferment for 5 days, turning the pile once a day.
[0084] (3) Seedling management
[0085] For the first two months of seedling cultivation, water once a day (with fresh water), and then once a week thereafter. When the seedlings are two months old, apply fertilizer solution once (5% NPK balanced compound fertilizer).
[0086] Comparative Example 1: A freshwater cultivation method for the mangrove plant *Gynostemma pentaphyllum*
[0087] In this embodiment, the amount of Veratrum nigrum leaf extract used is 50%.
[0088] (1) Seed treatment
[0089] Harvest mature female fruits of the sea olive (the pericarp changes from grayish-green to yellowish-green). Soak the harvested fruits in water for about 3 hours until the pericarp detaches from the seed (also known as the hypocotyl). Remove the pericarp floating on the surface and rinse the seeds 2-3 times with clean water. Soak the seeds in a 50% (w / w) veratrum leaf extract for 5 days.
[0090] Preparation method of Veratrum leaf extract: Take fresh Veratrum leaves, dry and pulverize them, extract with petroleum ether at 60℃ for 5 hours with stirring, filter, extract the filter residue with ethyl acetate at 70℃ for 10 hours, collect the ethyl acetate extract, and recover the ethyl acetate under reduced pressure to obtain Veratrum leaf extract.
[0091] Disinfect the female seeds of sea olives that have undergone soaking treatment by soaking them in a 500-fold dilution of chlorothalonil solution for 3 to 8 minutes, then remove and air dry them before sowing.
[0092] (2) Sowing
[0093] Seedling substrate: a mixture of sandy loam soil, dried cow manure, fermented material, and phosphate fertilizer (superphosphate, available phosphorus P2O5 ≥ 12%, calcium ≥ 16%), in a mass ratio of 80:2:10:10. Sow seeds in seedling bags containing the substrate, and place the seedling bags on ordinary sandy loam soil. Two weeks after sowing, count the germination rate (with radicle length greater than 3 mm as the germination indicator, n = 30).
[0094] The fermentation formula is as follows (parts by weight): 75 parts dried cow dung, 21 parts fan palm leaves, 1 part urea, and 2 parts Bacillus flexus bacterial solution. The preparation method involves taking dried cow dung, fan palm leaves, urea, and Bacillus flexus bacterial solution (containing 10% bacteria). 7 (cfu / mL), spread out, sprinkle with water and mix well (the amount of water should be such that when you squeeze the mixture in your hand, water drips out), pile it into a 1.0m×0.5m×0.2m long strip, ferment for 5 days, turning the pile once a day.
[0095] (3) Seedling management
[0096] For the first two months of seedling cultivation, water once a day (with fresh water), and then once a week thereafter. When the seedlings are two months old, apply fertilizer solution once (5% NPK balanced compound fertilizer).
[0097] Comparative Example 2: A freshwater cultivation method for the mangrove plant *Gynostemma pentaphyllum*
[0098] In this embodiment, the veratrum leaf extract was prepared by other methods.
[0099] (1) Seed treatment
[0100] Harvest mature female fruits of the sea olive (the pericarp changes from grayish-green to yellowish-green). Soak the harvested fruits in water for about 3 hours until the pericarp detaches from the seed (also known as the hypocotyl). Remove the pericarp floating on the surface and rinse the seeds 2-3 times with clean water. Soak the seeds in a 30% (w / w) veratrum leaf extract for 5 days.
[0101] Preparation method of Veratrum leaf extract: Take fresh Veratrum leaves, dry and pulverize them, extract with petroleum ether at 60℃ for 5 hours with stirring, filter, extract the filter residue with ethanol at 70℃ for 10 hours, collect the extract, and recover the ethanol under reduced pressure to obtain Veratrum leaf extract.
[0102] Disinfect the female seeds of sea olives that have undergone soaking treatment by soaking them in a 500-fold dilution of chlorothalonil solution for 3 to 8 minutes, then remove and air dry them before sowing.
[0103] (2) Sowing
[0104] Seedling substrate: a mixture of sandy loam soil, dried cow manure, fermented material, and phosphate fertilizer (superphosphate, available phosphorus P2O5 ≥ 12%, calcium ≥ 16%), in a mass ratio of 80:2:10:10. Sow seeds in seedling bags containing the substrate, and place the seedling bags on ordinary sandy loam soil. Two weeks after sowing, count the germination rate (with radicle length greater than 3 mm as the germination indicator, n = 30).
[0105] The fermentation formula is as follows (parts by weight): 75 parts dried cow dung, 21 parts fan palm leaves, 1 part urea, and 2 parts Bacillus flexus bacterial solution. The preparation method involves taking dried cow dung, fan palm leaves, urea, and Bacillus flexus bacterial solution (containing 10% bacteria). 7 (cfu / mL), spread out, sprinkle with water and mix well (the amount of water should be such that when you squeeze the mixture in your hand, water drips out), pile it into a 1.0m×0.5m×0.2m long strip, ferment for 5 days, turning the pile once a day.
[0106] (3) Seedling management
[0107] For the first two months of seedling cultivation, water once a day (with fresh water), and then once a week thereafter. When the seedlings are two months old, apply fertilizer solution once (5% NPK balanced compound fertilizer).
[0108] Comparative Example 3: A freshwater cultivation method for the mangrove plant *Gynostemma pentaphyllum*
[0109] In this embodiment, fermentation products prepared by other methods were selected.
[0110] (1) Seed treatment
[0111] Harvest mature female fruits of the sea olive (the pericarp changes from grayish-green to yellowish-green). Soak the harvested fruits in water for about 3 hours until the pericarp detaches from the seed (also known as the hypocotyl). Remove the pericarp floating on the surface and rinse the seeds 2-3 times with clean water. Soak the seeds in a 30% (w / w) veratrum leaf extract for 5 days.
[0112] Preparation method of Veratrum leaf extract: Take fresh Veratrum leaves, dry and pulverize them, extract with petroleum ether at 60℃ for 5 hours with stirring, filter, extract the filter residue with ethyl acetate at 70℃ for 10 hours, collect the ethyl acetate extract, and recover the ethyl acetate under reduced pressure to obtain Veratrum leaf extract.
[0113] Disinfect the female seeds of sea olives that have undergone soaking treatment by soaking them in a 500-fold dilution of chlorothalonil solution for 3 to 8 minutes, then remove and air dry them before sowing.
[0114] (2) Sowing
[0115] Seedling substrate: a mixture of sandy loam soil, dried cow manure, fermented material, and phosphate fertilizer (superphosphate, available phosphorus P2O5 ≥ 12%, calcium ≥ 16%), in a mass ratio of 80:2:10:10. Sow seeds in seedling bags containing the substrate, and place the seedling bags on ordinary sandy loam soil. Two weeks after sowing, count the germination rate (with radicle length greater than 3 mm as the germination indicator, n = 30).
[0116] The fermentation formula is as follows (parts by weight): 75 parts dried cow dung, 21 parts rice straw, 1 part urea, and 2 parts Bacillus flexus bacterial solution. The preparation method involves taking dried cow dung, rice straw, urea, and Bacillus flexus bacterial solution (containing 10% bacteria). 7 (cfu / mL), spread out, sprinkle with water and mix well (the amount of water should be such that when you squeeze the mixture in your hand, water drips out), pile it into a 1.0m×0.5m×0.2m long strip, ferment for 5 days, turning the pile once a day.
[0117] (3) Seedling management
[0118] For the first two months of seedling cultivation, water once a day (with fresh water), and then once a week thereafter. When the seedlings are two months old, apply fertilizer solution once (5% NPK balanced compound fertilizer).
[0119] Comparative Example 4: A freshwater cultivation method for the mangrove plant *Gynostemma pentaphyllum*
[0120] In this embodiment, fermentation products prepared by other methods were selected.
[0121] (1) Seed treatment
[0122] Harvest mature female fruits of the sea olive (the pericarp changes from grayish-green to yellowish-green). Soak the harvested fruits in water for about 3 hours until the pericarp detaches from the seed (also known as the hypocotyl). Remove the pericarp floating on the surface and rinse the seeds 2-3 times with clean water. Soak the seeds in a 30% (w / w) veratrum leaf extract for 5 days.
[0123] Preparation method of Veratrum leaf extract: Take fresh Veratrum leaves, dry and pulverize them, extract with petroleum ether at 60℃ for 5 hours with stirring, filter, extract the filter residue with ethyl acetate at 70℃ for 10 hours, collect the ethyl acetate extract, and recover the ethyl acetate under reduced pressure to obtain Veratrum leaf extract.
[0124] Disinfect the female seeds of sea olives that have undergone soaking treatment by soaking them in a 500-fold dilution of chlorothalonil solution for 3 to 8 minutes, then remove and air dry them before sowing.
[0125] (2) Sowing
[0126] Seedling substrate: a mixture of sandy loam soil, dried cow manure, fermented material, and phosphate fertilizer (superphosphate, available phosphorus P2O5 ≥ 12%, calcium ≥ 16%), in a mass ratio of 80:2:10:10. Sow seeds in seedling bags containing the substrate, and place the seedling bags on ordinary sandy loam soil. Two weeks after sowing, count the germination rate (with radicle length greater than 3 mm as the germination indicator, n = 30).
[0127] The fermentation formula is as follows (parts by weight): 75 parts dried cow dung, 21 parts fan palm leaves, 1 part urea, and 2 parts Bacillus subtilis inoculum. The preparation method involves taking dried cow dung, urea, fan palm leaves, and Bacillus subtilis inoculum (containing 10% bacteria). 7 (cfu / mL), spread out, sprinkle with water and mix well (the amount of water should be such that when you squeeze the mixture in your hand, water drips out), pile it into a 1.0m×0.5m×0.2m long strip, ferment for 5 days, turning the pile once a day.
[0128] (3) Seedling management
[0129] For the first two months of seedling cultivation, water once a day (with fresh water), and then once a week thereafter. When the seedlings are two months old, apply fertilizer solution once (5% NPK balanced compound fertilizer).
[0130] Comparative Example 5: A freshwater cultivation method for the mangrove plant *Gynostemma pentaphyllum*
[0131] In this embodiment, no fermentation material is used.
[0132] (1) Seed treatment
[0133] Harvest mature female fruits of the sea olive (the pericarp changes from grayish-green to yellowish-green). Soak the harvested fruits in water for about 3 hours until the pericarp detaches from the seed (also known as the hypocotyl). Remove the pericarp floating on the surface and rinse the seeds 2-3 times with clean water. Soak the seeds in a 30% (w / w) veratrum leaf extract for 5 days.
[0134] Preparation method of Veratrum leaf extract: Take fresh Veratrum leaves, dry and pulverize them, extract with petroleum ether at 60℃ for 5 hours with stirring, filter, extract the filter residue with ethyl acetate at 70℃ for 10 hours, collect the ethyl acetate extract, and recover the ethyl acetate under reduced pressure to obtain Veratrum leaf extract.
[0135] Disinfect the female seeds of sea olives that have undergone soaking treatment by soaking them in a 500-fold dilution of chlorothalonil solution for 3 to 8 minutes, then remove and air dry them before sowing.
[0136] (2) Sowing
[0137] Seedling substrate: a mixture of sandy loam soil, dried cow manure, and phosphate fertilizer (superphosphate, available phosphorus P2O5 ≥ 12%, calcium ≥ 16%), in a mass ratio of 80:2:10:10. Sow seeds in seedling bags containing the substrate, and place the bags on ordinary sandy loam soil. Two weeks after sowing, count the germination rate (with radicle length greater than 3 mm as the germination indicator, n = 30).
[0138] (3) Seedling management
[0139] For the first two months of seedling cultivation, water once a day (with fresh water), and then once a week thereafter. When the seedlings are two months old, apply fertilizer solution once (5% NPK balanced compound fertilizer).
[0140] Germination rate was recorded two weeks after sowing. Seedling height and survival rate were recorded six months after seedling cultivation. Each group underwent three parallel trials (n=30). Results are shown in Table 1.
[0141] Table 1
[0142] Germination rate / % Seedling height after 6 months of seedling cultivation / cm Survival rate / % Example 1 97.8 57.5 96.7 Example 2 96.7 55.2 94.4 Example 3 95.6 56.1 93.3 Example 4 96.7 55.1 93.3 Example 5 94.4 50.6 95.6 Comparative Example 1 80.0 45.0 88.9 Comparative Example 2 85.6 47.1 90.0 Comparative Example 3 97.8 43.8 86.7 Comparative Example 4 97.8 41.9 87.8 Comparative Example 5 97.8 41.4 83.3
[0143] Compared with the comparative example, the germination rate, seedling height and survival rate of the embodiment were significantly improved, with a germination rate of over 97%, a seedling height of 57cm after 6 months of seedling cultivation and a survival rate of over 96%.
[0144] The above description is only a partial embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.
Claims
1. A freshwater cultivation method for the mangrove plant *Gynostemma pentaphyllum*, characterized in that, Includes the following steps: (1) Seed treatment Take female seeds of sea olives and soak them in a seed soaking solution; after soaking, sterilize them and air dry them in the shade. The soaking solution is a 20-30% (w / w) extract of Veratrum nigrum leaves; The preparation method of the veratrum leaf extract includes: taking fresh veratrum leaves, drying and pulverizing them, extracting them with petroleum ether at 60~70℃ with stirring, filtering, extracting the filter residue with ethyl acetate at 70~75℃, collecting the ethyl acetate extract, and recovering the ethyl acetate under reduced pressure to obtain the veratrum leaf extract. (2) Sowing Prepare a seedling substrate and sow the seeds in a seedling bag containing the substrate; the seedling substrate contains sandy loam soil and poultry and livestock manure, fermented products and phosphate fertilizer in a mass ratio of 70~80:2~5:10~17:8~10. The fermentation formula, by weight, includes: 75-80 parts of poultry and livestock manure, 16.5-21 parts of palm leaves, 1-0.5 parts of urea, and 2-3 parts of Bacillus curvatureii bacterial solution; The method for making fermented products includes: mixing poultry and livestock manure, palm leaves, urea, and Bacillus flexuralis bacterial solution, spreading them out, sprinkling water and mixing them evenly, piling them into long strips, and fermenting for 3-5 days; (3) Seedling management Water regularly with fresh water and fertilize.
2. The freshwater cultivation method for the mangrove plant *Rhizophora stylosa* according to claim 1, characterized in that, The soaking time is 3-5 days.
3. The freshwater cultivation method for the mangrove plant *Rhizophora stylosa* according to claim 1, characterized in that, The preparation method of the veratrum leaf extract includes: taking fresh veratrum leaves, drying and pulverizing them, extracting them with petroleum ether at 60-70℃ with stirring for 5-7 hours, filtering, extracting the filter residue with ethyl acetate at 70-75℃ for 8-10 hours, collecting the ethyl acetate extract, and recovering the ethyl acetate under reduced pressure to obtain the veratrum leaf extract.
4. The freshwater cultivation method for the mangrove plant *Rhizophora stylosa* according to claim 1, characterized in that, The livestock manure in question is dried cow manure.
5. The freshwater cultivation method for the mangrove plant *Rhizophora stylosa* according to claim 1, characterized in that, The soaking solution was a 30% (w / w) extract of Veratrum nigrum leaves.
6. The freshwater cultivation method for the mangrove plant *Rhizophora stylosa* according to claim 1, characterized in that, The mass ratio of sandy loam soil to poultry and livestock manure, fermentation products, and phosphate fertilizer is 80:2:10:
10.
7. The freshwater cultivation method for the mangrove plant *Rhizophora stylosa* according to claim 1, characterized in that, The preparation method of Veratrum leaf extract includes: taking fresh Veratrum leaves, drying and pulverizing them, extracting them with petroleum ether at 60℃ for 5 hours with stirring, filtering, extracting the filter residue with ethyl acetate at 70℃ for 10 hours, collecting the ethyl acetate extract, and recovering the ethyl acetate under reduced pressure to obtain Veratrum leaf extract.
8. The freshwater cultivation method for the mangrove plant *Rhizophora stylosa* according to claim 1, characterized in that, The seedling management method is as follows: water with fresh water once a day for the first two months of seedling cultivation, and then water once a week thereafter; apply fertilizer water once when the seedlings are two months old.