A high yield rape cultivation method

Abstract

This invention relates to the field of rapeseed cultivation technology, specifically to a high-yield rapeseed cultivation method. The method includes the following steps: (1) preparation of TA / SA microspheres; (2) preparation of quercetin-platycodon polysaccharide; (3) seed pretreatment; (4) preparation of seedling substrate; (5) sowing; (6) seedling management; and (7) transplanting.

Description

Technical Field

[0001] This invention relates to the field of rapeseed cultivation technology, specifically to a high-yield rapeseed cultivation method. Background Technology

[0002] Rapeseed is an important oilseed crop, with oil accounting for 35-50% of its dry weight. Rapeseed oil is a high-quality edible vegetable oil, rich in fatty acids and various vitamins, and highly nutritious. The oil cake after pressing contains about 40% protein, with a nutritional value similar to soybean cake, making it a good concentrated feed. Returning residual roots, stems, and shells to the field can significantly improve soil fertility. Current rapeseed cultivation techniques mainly rely on a large number of seedlings to increase yield, which suffers from problems such as high seed consumption, significant fertilizer loss, unstable yields, and low economic benefits. Direct-seeded rapeseed has low germination rates and poor uniformity; low fertilizer utilization and severe nutrient loss; and excessive application of chemical fertilizers leads to soil compaction. Therefore, developing a high-yield rapeseed cultivation method is of great significance for improving the economic benefits of rapeseed planting. Summary of the Invention

[0003] To address the shortcomings of existing technologies, this invention proposes a high-yield rapeseed cultivation method.

[0004] This invention is achieved through the following technical solution: A high-yield rapeseed cultivation method includes the following steps: (1) Preparation of TA / SA microspheres: (11) Dissolve tannic acid (TA) in deionized water, add sodium alginate (SA) and stir to dissolve to obtain a mixed solution. Add the mixed solution dropwise into 3-5wt% CaCl2 solution, solidify for 12 h, filter, wash with deionized water, freeze dry to obtain TA / SA microspheres. (12) The TA / SA microspheres obtained in step (11) were mixed with epichlorohydrin, stirred at 80°C, and 10% NaOH aqueous solution was added dropwise. The mixture was stirred at 80°C for 3 h, cooled to room temperature, filtered, washed with ethanol and deionized water, and dried under vacuum to obtain epoxy TA / SA microspheres. (13) Add the epoxy TA / SA microspheres obtained in step (12) and sodium carbonate to deionized water, add triethylenetetramine dropwise under stirring, stir at 50°C for 3-4 h, filter, wash with ethanol and deionized water, and vacuum dry to obtain modified TA / SA microspheres. (2) Preparation of quercetin-Platycodon grandiflorum polysaccharide: (21) Wash and dry the Platycodon grandiflorus, crush it through a 60-80 mesh sieve, add it to an 80% v / v ethanol aqueous solution at a material-to-liquid ratio of 1 g: 20 mL, soak it for 12 h, sonicate it at 50℃ and 600-800W for 2 h, filter it, dry the filter residue, and obtain the pretreated Platycodon grandiflorus. (22) The pretreated Platycodon grandiflorus obtained in step (21) was added to distilled water at a material-to-liquid ratio of 1 g: 20-30 mL, and extracted at 90℃ for 2 h. The extraction was repeated 3 times. The extracts were combined and concentrated to 1 / 5 of the original volume. Four times the volume of ethanol was added, and the mixture was allowed to stand at 4℃ for 12-16 h. The mixture was centrifuged at 5000-6000 rpm for 10 min. The precipitate was eluted with anhydrous ether and acetone and filtered. The protein was removed by trichloroacetic acid. The mixture was dialyzed in ultrapure water for 72 h and then freeze-dried to obtain Platycodon grandiflorus polysaccharide. (23) Take the Platycodon grandiflorum polysaccharide, TEMPO and NaBr obtained in step (22) and add them to ultrapure water. Stir to dissolve. Add NaClO solution with 5-10% active chlorine content. Maintain the pH value at 10-11 with NaOH solution. Stir for 4 h. Add ethanol and stir for 5 min. Adjust the pH to 7 with HCl solution. Add 5 times the volume of ethanol. Let stand for 2 h. Centrifuge at 8000 r / min for 10-15 min. Wash the precipitate with ethanol. Dialyze with ultrapure water for 72 h. Freeze dry to obtain oxidized Platycodon grandiflorum polysaccharide. (24) Dissolve the oxidized Platycodon polysaccharide obtained in step (23) in PBS solution at pH 5.5, add EDC·HCl and NHS, stir for 1 h, add 3-aminophenylboronic acid, react for 24 h under nitrogen protection, dialyze with deionized water for 3 days, freeze dry to obtain Platycodon polysaccharide-PBA; (25) Add quercetin to ethanol and sonicate to dissolve it to obtain a quercetin solution. Dissolve the platycodon polysaccharide-PBA obtained in step (24) in a PBS solution with pH 7.4, add the quercetin solution, sonicate to disperse, stir at 500-600 rpm for 2-3 h in the dark at 60℃, dialyze to a PBS solution with pH 7.4 for 24 h, dialyze to ultrapure water for 6 h, and freeze dry to obtain quercetin-platycodon polysaccharide. (3) Seed pretreatment: Add quercetin-placos polysaccharide to ultrapure water at a concentration of 0.3-0.5 g / L and mix well to obtain quercetin-placos polysaccharide solution. Select rapeseed seeds that are uniform in size, plump and undamaged. After disinfection, soak the seeds in the quercetin-placos polysaccharide solution for 18-24 h, take them out and air dry. (4) Preparation of seedling substrate: Mix peat moss, perlite and vermiculite in a volume ratio of 6:3:1 to obtain the basic substrate. Add 10-15 g / kg of decomposed organic fertilizer and 5-8 g / kg of NPK compound fertilizer (15:15:15) to obtain the seedling substrate. Adjust the substrate moisture content to 60%-70%. (5) Sowing: After sterilizing the seedling substrate, fill it into the seedling tray. Sow 2 seeds per hole after the pretreatment in step (3), at a depth of 1.0-1.5 cm. Cover with substrate 0.5 cm and water thoroughly. (6) Seedling management: After sowing, maintain a daytime temperature of 20-25℃ and a nighttime temperature of 12-15℃. When the seedlings have grown two true leaves, thin them out, leaving one seedling per hole. Cultivate the seedlings until they have 5-6 true leaves, then they are ready for transplanting. (7) Transplanting: Transplant when the soil temperature is stable above 5℃ after the soil thaws in spring. Apply 30-40 kg of NPK compound fertilizer and 1.0-1.5 kg of borax per mu. At the same time, apply 8-12 kg / mu of modified TA / SA microspheres. Spread evenly and then plow into the soil to a depth of 20-25 cm. When lifting the seedlings, keep the intact substrate soil ball and plant them at a spacing of 20 cm × 40 cm. Water thoroughly to settle the roots. After planting, carry out routine field management.

[0005] Further, in step (11), the mass concentration of tannic acid in deionized water is 30-40 mg / mL.

[0006] Further, in step (11), the mass ratio of tannic acid to sodium alginate is 2:1.

[0007] Further, in step (11), the mass ratio of sodium alginate to CaCl2 is 1:2.

[0008] Further, in step (12), the mass ratio of the TA / SA microspheres, epichlorohydrin and NaOH aqueous solution is 1.5:3:2.

[0009] Further, in step (13), the mass concentration of the epoxy TA / SA microspheres in deionized water is 20 mg / mL.

[0010] Further, in step (13), the mass ratio of the epoxy TA / SA microspheres, sodium carbonate and triethylenetetramine is 4:1:3.

[0011] Further, in step (23), the mass concentration of the platycodon polysaccharide in ultrapure water is 5-10 mg / mL.

[0012] Further, in step (23), the ratio of the amount of Platycodon grandiflorum polysaccharide, TEMPO, NaBr and NaClO solution is 1 g:0.02 g:0.2 g:5 mL.

[0013] Furthermore, in step (23), the volume ratio of ethanol to ultrapure water is 1:10.

[0014] Further, in step (24), the mass concentration of the oxidized platycodon polysaccharide in the PBS solution is 10-20 mg / mL.

[0015] Further, in step (24), the mass ratio of the oxidized platycodon polysaccharide, 3-aminophenylboronic acid, EDC·HCl and NHS is 1:0.3:0.5:0.3.

[0016] Further, in step (25), the mass concentration of quercetin in ethanol is 2 mg / mL.

[0017] Further, in step (25), the mass concentration of the platycodon polysaccharide-PBA in the PBS solution is 2 mg / mL.

[0018] Further, in step (25), the mass ratio of quercetin to platycodon polysaccharide-PBA is 1:4.

[0019] Compared with the prior art, the present invention has the following beneficial effects: This invention provides a high-yield rapeseed cultivation method that significantly improves seed germination rate and increases yield.

[0020] This invention prepares a quercetin-placose polysaccharide. First, a carboxyl group is introduced onto the platycose polysaccharide molecule via a TEMPO oxidation system. Then, 3-aminophenylboronic acid is grafted onto the polysaccharide using EDC / NHS catalysis. Finally, the borate group in the PBA molecule forms a borate ester bond with the ortho-dihydroxy group of quercetin, reversibly covalently linking quercetin and platycose polysaccharide through 3-aminophenylboronic acid. Using platycose polysaccharide, which has good water solubility and high biocompatibility, as a carrier, quercetin is loaded onto the polysaccharide molecule, significantly improving the dispersibility and cell membrane permeability of quercetin, allowing it to enter the seed interior more efficiently and exert its effects. As a plant-derived active polysaccharide, platycose polysaccharide can provide sufficient energy and material basis for embryo growth by regulating seed cell membrane permeability. Quercetin, as a strong antioxidant among natural flavonoids, can effectively eliminate excess reactive oxygen species generated during seed germination due to increased respiration. It can also regulate the balance of endogenous hormones in seeds, break seed dormancy, and significantly improve germination potential and germination rate. This invention employs quercetin-platycodon polysaccharide pretreatment of rapeseed. Quercetin-platycodon polysaccharide possesses antioxidant activity, biocompatibility, and cell metabolism-promoting effects. The combination of these two substances effectively enhances seed vigor, increases germination potential, and improves germination rate. This invention also prepares TA / SA microspheres, which are cross-linked with sodium alginate and tannic acid to form a three-dimensional interconnected network structure, increasing specific surface area, reducing soil moisture evaporation and nutrient leaching, and improving water and fertilizer utilization. Furthermore, this invention introduces epoxy groups through the reaction of epichlorohydrin with the phenolic hydroxyl groups of tannic acid, followed by a ring-opening reaction with triethylenetetramine, introducing amino groups onto the surface and interior of the microspheres. These groups can bind negatively charged anionic nutrients in the soil through electrostatic adsorption. The microspheres can also slowly biodegrade, gradually releasing nutrients. Tannic acid molecules contain a large number of phenolic hydroxyl groups, which can interact with humus in the soil through hydrogen bonds and coordination bonds; the amino groups on the surface of modified TA / SA microspheres can enhance the cementation effect on soil particles, combine with the negative charge on the surface of soil particles, act as a bridge between soil particles, and promote the formation of large soil aggregates; the microspheres themselves are natural biodegradable polymer materials, and the degradation products are organic matter and humus precursors, which can further increase the soil organic matter content and improve soil aeration and permeability. Attached Figure Description

[0021] Figure 1 The germination rate of the seed pretreatment methods described in Examples 1-3 and Comparative Examples 1-3 of this invention; Figure 2 The yields of the cultivation methods described in Examples 1-3 and Comparative Examples 1-5 of this invention; Figure 3 The soil improvement effect of the transplanting methods described in Examples 1-3 and Comparative Examples 4-5 of this invention. Detailed Implementation

[0022] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to specific embodiments. However, this invention is not limited to the following embodiments. It should be noted that, unless otherwise specified, all chemical reagents involved in this invention are purchased through commercial channels.

[0023] Example 1: A high-yield rapeseed cultivation method, comprising the following steps: (1) Preparation of TA / SA microspheres: (11) Take 2 g of tannic acid (TA) and dissolve it in 50 mL of deionized water. Add 1 g of sodium alginate (SA) and stir to dissolve it to obtain a mixed solution. Add the mixed solution dropwise into a 5 wt% CaCl2 solution containing 2 g CaCl2, solidify for 12 h, filter, wash with deionized water, freeze dry to obtain TA / SA microspheres. (12) Mix 3 g of TA / SA microspheres obtained in step (11) with 6 g of epichlorohydrin, stir at 80°C, add 4 g of 10% NaOH aqueous solution, stir at 80°C for 3 h, cool to room temperature, filter, wash with ethanol and deionized water, and vacuum dry to obtain epoxy TA / SA microspheres. (13) Add 2 g of epoxy TA / SA microspheres obtained in step (12) and 0.5 g of sodium carbonate to 100 mL of deionized water, add 1.5 g of triethylenetetramine dropwise under stirring, stir at 50 °C for 4 h, filter, wash with ethanol and deionized water, and vacuum dry to obtain modified TA / SA microspheres. (2) Preparation of quercetin-Platycodon grandiflorum polysaccharide: (21) Wash and dry the Platycodon grandiflorus, crush it through an 80-mesh sieve, add it to an 80% v / v ethanol aqueous solution at a material-to-liquid ratio of 1 g: 20 mL, soak it for 12 h, sonicate it at 50℃ and 800W for 2 h, filter it, dry the filter residue, and obtain the pretreated Platycodon grandiflorus. (22) The pretreated Platycodon grandiflorus obtained in step (21) was added to distilled water at a material-to-liquid ratio of 1 g: 30 mL, and extracted by heating at 90°C for 2 h. The extraction was repeated 3 times. The extracts were combined and concentrated to 1 / 5 of the original volume. Four times the volume of ethanol was added, and the mixture was allowed to stand at 4°C for 16 h. The mixture was centrifuged at 6000 rpm for 10 min. The precipitate was eluted with anhydrous ether and acetone and filtered. The protein was removed by trichloroacetic acid. The mixture was dialyzed in ultrapure water for 72 h and then freeze-dried to obtain Platycodon grandiflorus polysaccharide. (23) Take 1 g of Platycodon grandiflorum polysaccharide obtained in step (22), 0.02 g of TEMPO and 0.2 g of NaBr and add them to 100 mL of ultrapure water. Stir to dissolve, add 5 mL of NaClO solution with 10% active chlorine content, maintain the pH value at 11 with NaOH solution, stir for 4 h, add 10 mL of ethanol, stir for 5 min, adjust the pH to 7 with HCl solution, add 5 times the volume of ethanol, let stand for 2 h, centrifuge at 8000 r / min for 15 min, wash the precipitate with ethanol, dialyze with ultrapure water for 72 h, freeze dry to obtain oxidized Platycodon grandiflorum polysaccharide; (24) Take 1 g of the oxidized Platycodon polysaccharide obtained in step (23) and dissolve it in 50 mL of PBS solution at pH 5.5. Add 0.5 g of EDC·HCl and 0.3 g of NHS, stir for 1 h, add 0.3 g of 3-aminophenylboronic acid, react under nitrogen protection for 24 h, dialyze with deionized water for 3 days, freeze dry to obtain Platycodon polysaccharide-PBA; (25) Add 0.25 g of quercetin to 125 mL of ethanol and sonicate to dissolve to obtain quercetin solution. Dissolve 1 g of Platycodon grandiflorum polysaccharide-PBA obtained in step (24) in 500 mL of PBS solution with pH 7.4, add quercetin solution, sonicate to disperse, stir at 600 rpm for 3 h in the dark at 60°C, dialyze to PBS solution with pH 7.4 for 24 h, dialyze to ultrapure water for 6 h, freeze dry to obtain quercetin-Platycodon grandiflorum polysaccharide; (3) Seed pretreatment: Quercetin-Platycodon polysaccharide was added to ultrapure water at a concentration of 0.5 g / L and mixed well to obtain quercetin-Platycodon polysaccharide solution. Spring rapeseed seeds of uniform size, plump and undamaged were selected, and after disinfection, they were soaked in quercetin-Platycodon polysaccharide solution for 24 h, and then taken out and dried. (4) Preparation of seedling substrate: Mix peat moss, perlite and vermiculite in a volume ratio of 6:3:1 to obtain the basic substrate. Add 15 g / kg of decomposed organic fertilizer and 8 g / kg of NPK compound fertilizer (15:15:15) to obtain the seedling substrate. Adjust the substrate moisture content to 70%. (5) Sowing: After sterilizing the seedling substrate, fill it into the seedling tray. Take the seeds that have been pretreated in step (3), sow 2 seeds per hole at a depth of 1.5 cm, cover with 0.5 cm of substrate, and water thoroughly. (6) Seedling management: After sowing, maintain a daytime temperature of 25℃ and a nighttime temperature of 15℃. When the seedlings have grown two true leaves, thin them out, leaving one seedling per hole. Cultivate them until they have six true leaves, then they are ready for transplanting. (7) Transplanting: Transplant when the soil temperature is stable above 5℃ after the soil thaws in spring. Apply 40 kg of nitrogen, phosphorus and potassium compound fertilizer and 1.5 kg of borax per mu. At the same time, apply 12 kg / mu of modified TA / SA microspheres. Spread evenly and then plow into the soil to a depth of 25 cm. When lifting the seedlings, keep the intact substrate soil ball and plant them at a spacing of 20 cm × 40 cm. Water thoroughly to settle the roots. After planting, carry out routine field management.

[0024] Example 2: A high-yield rapeseed cultivation method, comprising the following steps: (1) Preparation of TA / SA microspheres: (11) Take 2.4 g of tannic acid (TA) and dissolve it in 80 mL of deionized water. Add 1.2 g of sodium alginate (SA) and stir to dissolve it to obtain a mixed solution. Add the mixed solution dropwise into a 3wt% CaCl2 solution containing 2.4 g of CaCl2, solidify for 12 h, filter, wash with deionized water, freeze dry to obtain TA / SA microspheres; (12) Mix 3 g of TA / SA microspheres obtained in step (11) with 6 g of epichlorohydrin, stir at 80°C, add 4 g of 10% NaOH aqueous solution, stir at 80°C for 3 h, cool to room temperature, filter, wash with ethanol and deionized water, and vacuum dry to obtain epoxy TA / SA microspheres. (13) Add 2 g of epoxy TA / SA microspheres obtained in step (12) and 0.5 g of sodium carbonate to 100 mL of deionized water, add 1.5 g of triethylenetetramine dropwise under stirring, stir at 50 °C for 3 h, filter, wash with ethanol and deionized water, and vacuum dry to obtain modified TA / SA microspheres. (2) Preparation of quercetin-Platycodon grandiflorum polysaccharide: (21) Wash and dry the Platycodon grandiflorus, crush it through a 60-mesh sieve, add it to an 80% v / v ethanol aqueous solution at a material-to-liquid ratio of 1 g: 20 mL, soak it for 12 h, sonicate it at 50℃ and 600W for 2 h, filter it, dry the filter residue, and obtain the pretreated Platycodon grandiflorus. (22) The pretreated Platycodon grandiflorus obtained in step (21) was added to distilled water at a material-to-liquid ratio of 1 g: 20 mL, and extracted by heating at 90°C for 2 h. The extraction was repeated 3 times. The extracts were combined and concentrated to 1 / 5 of the original volume. Four times the volume of ethanol was added, and the mixture was allowed to stand at 4°C for 12 h. The mixture was centrifuged at 5000 rpm for 10 min. The precipitate was eluted with anhydrous ether and acetone and filtered. The protein was removed by trichloroacetic acid. The mixture was dialyzed in ultrapure water for 72 h and then freeze-dried to obtain Platycodon grandiflorus polysaccharide. (23) Take 1 g of Platycodon grandiflorum polysaccharide obtained in step (22), 0.02 g of TEMPO and 0.2 g of NaBr and add them to 200 mL of ultrapure water. Stir to dissolve, add 5 mL of NaClO solution with 5% active chlorine content, maintain the pH value at 10 with NaOH solution, stir for 4 h, add 20 mL of ethanol, stir for 5 min, adjust the pH to 7 with HCl solution, add 5 times the volume of ethanol, let stand for 2 h, centrifuge at 8000 r / min for 10 min, wash the precipitate with ethanol, dialyze with ultrapure water for 72 h, freeze dry to obtain oxidized Platycodon grandiflorum polysaccharide; (24) Take 1 g of the oxidized Platycodon grandiflorum polysaccharide obtained in step (23) and dissolve it in 100 mL of PBS solution at pH 5.5. Add 0.5 g of EDC·HCl and 0.3 g of NHS, stir for 1 h, add 0.3 g of 3-aminophenylboronic acid, react under nitrogen protection for 24 h, dialyze with deionized water for 3 days, freeze dry to obtain Platycodon grandiflorum polysaccharide-PBA; (25) Add 0.25 g of quercetin to 125 mL of ethanol and sonicate to dissolve to obtain quercetin solution. Dissolve 1 g of Platycodon grandiflorum polysaccharide-PBA obtained in step (24) in 500 mL of PBS solution with pH 7.4, add quercetin solution, sonicate to disperse, stir at 500 rpm for 2 h in the dark at 60℃, dialyze to PBS solution with pH 7.4 for 24 h, dialyze to ultrapure water for 6 h, freeze dry to obtain quercetin-Platycodon grandiflorum polysaccharide. (3) Seed pretreatment: Quercetin-Platycodon polysaccharide was added to ultrapure water at a concentration of 0.3 g / L and mixed well to obtain quercetin-Platycodon polysaccharide solution. Rapeseeds of uniform size, plump and undamaged were selected, disinfected and soaked in quercetin-Platycodon polysaccharide solution for 18 h, and then taken out and dried. (4) Preparation of seedling substrate: Mix peat moss, perlite and vermiculite in a volume ratio of 6:3:1 to obtain the basic substrate. Add 10 g / kg of decomposed organic fertilizer and 5 g / kg of NPK compound fertilizer (15:15:15) to obtain the seedling substrate. Adjust the substrate moisture content to 60%. (5) Sowing: After sterilizing the seedling substrate, fill it into the seedling tray. Sow 2 seeds per hole after the pretreatment in step (3), at a depth of 1.0 cm. Cover with substrate 0.5 cm and water thoroughly. (6) Seedling management: After sowing, maintain a daytime temperature of 20℃ and a nighttime temperature of 12℃. When the seedlings have grown two true leaves, thin them out, leaving one seedling per hole. Cultivate them until they have five true leaves, and then they are ready for transplanting. (7) Transplanting: Transplant when the soil temperature is stable above 5℃ after the soil thaws in spring. Apply 30 kg of nitrogen, phosphorus and potassium compound fertilizer and 1.0 kg of borax per mu. At the same time, apply 8 kg / mu of modified TA / SA microspheres. Spread evenly and then plow into the soil to a depth of 20 cm. When lifting the seedlings, keep the intact substrate soil ball and plant them at a spacing of 20 cm × 40 cm. Water thoroughly to settle the roots. After planting, carry out routine field management.

[0025] Example 3: A high-yield rapeseed cultivation method, comprising the following steps: (1) Preparation of TA / SA microspheres: (11) Take 2 g of tannic acid (TA) and dissolve it in 60 mL of deionized water. Add 1 g of sodium alginate (SA) and stir to dissolve it to obtain a mixed solution. Add the mixed solution dropwise into a 4 wt% CaCl2 solution containing 2 g CaCl2, solidify for 12 h, filter, wash with deionized water, freeze dry to obtain TA / SA microspheres. (12) Mix 3 g of TA / SA microspheres obtained in step (11) with 6 g of epichlorohydrin, stir at 80°C, add 4 g of 10% NaOH aqueous solution, stir at 80°C for 3 h, cool to room temperature, filter, wash with ethanol and deionized water, and vacuum dry to obtain epoxy TA / SA microspheres. (13) Add 2 g of epoxy TA / SA microspheres obtained in step (12) and 0.5 g of sodium carbonate to 100 mL of deionized water, add 1.5 g of triethylenetetramine dropwise under stirring, stir at 50 °C for 3-4 h, filter, wash with ethanol and deionized water, and vacuum dry to obtain modified TA / SA microspheres. (2) Preparation of quercetin-Platycodon grandiflorum polysaccharide: (21) Wash and dry the Platycodon grandiflorus, crush it through a 70-mesh sieve, add it to an 80% v / v ethanol aqueous solution at a material-to-liquid ratio of 1 g: 20 mL, soak it for 12 h, sonicate it at 50℃ and 700W for 2 h, filter it, dry the filter residue, and obtain the pretreated Platycodon grandiflorus. (22) The pretreated Platycodon grandiflorus obtained in step (21) was added to distilled water at a material-to-liquid ratio of 1 g: 25 mL, and extracted at 90°C for 2 h. The extraction was repeated 3 times. The extracts were combined and concentrated to 1 / 5 of the original volume. Four times the volume of ethanol was added, and the mixture was allowed to stand at 4°C for 14 h. The mixture was centrifuged at 5500 rpm for 10 min. The precipitate was eluted with anhydrous ether and acetone and filtered. The protein was removed by trichloroacetic acid. The mixture was dialyzed in ultrapure water for 72 h and then freeze-dried to obtain Platycodon grandiflorus polysaccharide. (23) Take 1 g of Platycodon grandiflorum polysaccharide obtained in step (22), 0.02 g of TEMPO and 0.2 g of NaBr and add them to 150 mL of ultrapure water. Stir to dissolve, add 5 mL of NaClO solution with 8% active chlorine content, maintain the pH value at 10.8 with NaOH solution, stir for 4 h, add 15 mL of ethanol, stir for 5 min, adjust the pH to 7 with HCl solution, add 5 times the volume of ethanol, let stand for 2 h, centrifuge at 8000 r / min for 12 min, wash the precipitate with ethanol, dialyze with ultrapure water for 72 h, freeze dry to obtain oxidized Platycodon grandiflorum polysaccharide; (24) Take 1 g of the oxidized Platycodon grandiflorum polysaccharide obtained in step (23) and dissolve it in 80 mL of PBS solution at pH 5.5. Add 0.5 g of EDC·HCl and 0.3 g of NHS, stir for 1 h, add 0.3 g of 3-aminophenylboronic acid, react under nitrogen protection for 24 h, dialyze with deionized water for 3 days, freeze dry to obtain Platycodon grandiflorum polysaccharide-PBA; (25) Add 0.25 g of quercetin to 125 mL of ethanol and sonicate to dissolve to obtain quercetin solution. Dissolve 1 g of Platycodon grandiflorum polysaccharide-PBA obtained in step (24) in 500 mL of PBS solution with pH 7.4, add quercetin solution, sonicate to disperse, stir at 550 rpm for 2.5 h in the dark at 60℃, dialyze to PBS solution with pH 7.4 for 24 h, dialyze to ultrapure water for 6 h, freeze dry to obtain quercetin-Platycodon grandiflorum polysaccharide; (3) Seed pretreatment: Quercetin-Platycodon polysaccharide was added to ultrapure water at a concentration of 0.4 g / L and mixed well to obtain quercetin-Platycodon polysaccharide solution. Rapeseeds of uniform size, plump and undamaged were selected, disinfected and soaked in quercetin-Platycodon polysaccharide solution for 20 h, and then taken out and dried. (4) Preparation of seedling substrate: Mix peat moss, perlite and vermiculite in a volume ratio of 6:3:1 to obtain the basic substrate. Add 12 g / kg of decomposed organic fertilizer and 6 g / kg of NPK compound fertilizer (15:15:15) to obtain the seedling substrate. Adjust the substrate moisture content to 65%. (5) Sowing: After sterilizing the seedling substrate, fill it into the seedling trays. Sow 2 seeds per hole after the pretreatment in step (3) at a depth of 1.2 cm. Cover with substrate 0.5 cm and water thoroughly. (6) Seedling management: After sowing, maintain a daytime temperature of 22℃ and a nighttime temperature of 14℃. When the seedlings have grown two true leaves, thin them out, leaving one seedling per hole. Cultivate them until they have six true leaves, then they are ready for transplanting. (7) Transplanting: Transplant when the soil temperature is stable above 5℃ after the soil thaws in spring. Apply 35 kg of nitrogen, phosphorus and potassium compound fertilizer and 1.2 kg of borax per mu. At the same time, apply 10 kg / mu of modified TA / SA microspheres. Spread evenly and then plow into the soil to a depth of 22 cm. When lifting the seedlings, keep the intact substrate soil ball and plant them at a spacing of 20 cm × 40 cm. Water thoroughly to settle the roots. After planting, carry out routine field management.

[0026] The only difference between Comparative Example 1 and Example 1 is that Platycodon grandiflorum polysaccharide is used instead of quercetin-Platycodon grandiflorum polysaccharide.

[0027] The only difference between Comparative Example 2 and Example 1 is that quercetin is used instead of quercetin-platycodon polysaccharide.

[0028] The only difference between Comparative Example 3 and Example 1 is that quercetin-placosaccharide was not added.

[0029] The only difference between Comparative Example 4 and Example 1 is that TA / SA microspheres were used instead of modified TA / SA microspheres.

[0030] The only difference between Comparative Example 5 and Example 1 is that no modified TA / SA microspheres were added.

[0031] Experimental Example 1: Following the methods of Examples 1-3 and Comparative Examples 1-3, spring rapeseed seeds were pretreated. The treated seeds were placed in a petri dish lined with double layers of moist filter paper, and covered with a moist, clean gauze. During cultivation, the seeds were watered every 12 hours (enough to moisten the filter paper). Germination was initiated in a 25℃ constant temperature incubator. The number of germinations was recorded daily (based on the seed radicle breaking through the seed coat by ≥2 mm). Germination potential was calculated on the 3rd day, and the germination rate was calculated on the 7th day. Results are as follows: Figure 1 As shown.

[0032] Figure 1 The results showed that the germination rate and germination potential of Examples 1-3 were significantly higher than those of Comparative Examples 1-3. Comparative Example 1 used platycodon polysaccharide instead of quercetin-platycodon polysaccharide, and Comparative Example 2 used quercetin instead of quercetin-platycodon polysaccharide, resulting in a decreased effect on promoting seed germination. Comparative Example 3 did not add quercetin-platycodon polysaccharide, but only soaked in water; its germination rate was lower than that of Comparative Examples 1-3 and Comparative Examples 1-2. These results indicate that the pretreatment method for rapeseed seeds of the present invention has a significant promoting effect on seed germination, can improve seed stress resistance, and increase yield.

[0033] Experimental Example 2: Rapeseed was cultivated according to the methods of Examples 1-3 and Comparative Examples 1-5. Transplanting was done in the same plot of land, following the methods of Examples 1-3 and Comparative Examples 1-5. Yield was measured before harvest after maturity, and the results are as follows: Figure 2 As shown.

[0034] Figure 2The results showed that the yields of Examples 1-3 were significantly higher than those of Comparative Examples 1-5. Comparative Example 1 used Platycodon grandiflorum polysaccharide instead of quercetin-Platycodon grandiflorum polysaccharide; Comparative Example 2 used quercetin instead of quercetin-Platycodon grandiflorum polysaccharide; Comparative Example 3 did not add quercetin-Platycodon grandiflorum polysaccharide; Comparative Example 4 used TA / SA microspheres instead of modified TA / SA microspheres; and Comparative Example 5 and the group without modified TA / SA microspheres all showed reduced yields. This indicates that the rapeseed cultivation method of the present invention can effectively increase yield.

[0035] Experimental Example 3: Soil samples from the 0-20 cm topsoil layer were collected in the experimental areas of Examples 1-3 and Comparative Examples 4-5 after rapeseed harvest. Soil aggregate composition was determined, and the content of large aggregates (>0.25 mm) was calculated. Results are as follows: Figure 3 As shown.

[0036] Figure 3 The results showed that the content of large aggregates in Examples 1-3 was significantly higher than that in Comparative Examples 4-5. Comparative Example 4 used TA / SA microspheres instead of modified TA / SA microspheres, while Comparative Example 5 did not add modified TA / SA microspheres, resulting in a decreased soil structure improvement effect. The rapeseed cultivation method of this invention can effectively improve soil and increase yield.

[0037] 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 high yield oilseed rape cultivation method, characterized in that, Includes the following steps: (1) Preparation of TA / SA microspheres: (11) Dissolve tannic acid and sodium alginate in water, add them dropwise into CaCl2 solution, and solidify to obtain TA / SA microspheres; (12) The TA / SA microspheres obtained in step (11) are reacted with epichlorohydrin to obtain epoxy-based TA / SA microspheres; (13) Add the epoxy TA / SA microspheres obtained in step (12) and sodium carbonate to deionized water, add triethylenetetramine, stir and react to obtain modified TA / SA microspheres. (2) Preparation of quercetin-Platycodon grandiflorum polysaccharide: (21) Platycodon grandiflorus was soaked in an ethanol aqueous solution to obtain pretreated Platycodon grandiflorus; (22) Add the pretreated Platycodon grandiflorus obtained in step (21) to distilled water, heat and extract, concentrate the extract, add ethanol, let stand, centrifuge, elute the precipitate with anhydrous ether and acetone, deproteinize with trichloroacetic acid, dialyze, freeze dry, and obtain Platycodon grandiflorus polysaccharide. (23) Take the Platycodon grandiflorum polysaccharide obtained in step (22) and oxidize it with TEMPO, NaBr and NaClO solution with active chlorine content of 5-10% to obtain oxidized Platycodon grandiflorum polysaccharide; (24) Dissolve the oxidized Platycodon polysaccharide obtained in step (23) in PBS solution, add EDC·HCl and NHS, stir, add 3-aminophenylboronic acid, react, dialyze, freeze dry to obtain Platycodon polysaccharide-PBA; (25) Quercetin was added to ethanol to obtain a quercetin solution. Platycodon polysaccharide-PBA obtained in step (24) was dissolved in PBS solution, and the quercetin solution was added. The mixture was dispersed, stirred, dialyzed, and freeze-dried to obtain quercetin-Platycodon polysaccharide. (3) Seed pretreatment: Quercetin-Platycodon polysaccharide was added to ultrapure water and mixed well, and then rapeseed seeds were soaked in the solution. (4) Preparation of seedling substrate: Take peat moss, perlite and vermiculite, mix them evenly, add fertilizer to obtain seedling substrate; (5) Sowing: Fill the seedling substrate into the seedling tray and sow the seeds after pretreatment in step (3); (6) Seedling management: Thin out seedlings, cultivate them until they have 5-6 true leaves, and then transplant them; (7) Transplanting: Apply soil fertilizer, apply modified TA / SA microspheres, plant, and perform routine field management.

2. The high yield oilseed rape cultivation method according to claim 1, characterized in that, In step (11), the mass ratio of tannic acid to sodium alginate is 2:

1.

3. The high yield oilseed rape cultivation method according to claim 2, characterized in that, In step (12), the mass ratio of TA / SA microspheres to epichlorohydrin is 1.5:

3.

4. The high-yield rapeseed cultivation method according to claim 3, characterized in that, In step (13), the mass ratio of the epoxy-based TA / SA microspheres to triethylenetetramine is 4:

3.

5. The high-yield rapeseed cultivation method according to claim 4, characterized in that, In step (23), the ratio of the amount of Platycodon grandiflorum polysaccharide, TEMPO, NaBr and NaClO solution is 1 g:0.02 g:0.2 g:5 mL.

6. The high-yield rapeseed cultivation method according to claim 5, characterized in that, In step (24), the mass ratio of the oxidized platycodon polysaccharide, 3-aminophenylboronic acid, EDC·HCl and NHS is 1:0.3:0.5:0.

3.

7. The high-yield rapeseed cultivation method according to claim 6, characterized in that, In step (25), the mass ratio of quercetin to Platycodon grandiflorum polysaccharide-PBA is 1:4.

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