Application of natural polysaccharide in preparation of seed initiator, plant growth regulator and / or resistance inducer
Chitosan application addresses soil and climate challenges in watermelon cultivation by improving seed germination, plant growth, and disease resistance, enhancing yield and quality.
Patent Information
- Application Number
- CN202510440072.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-09
- Publication Date
- 2025-07-15
AI Technical Summary
Watermelon production faces the problems of decline in yield and quality caused by factors such as soil conditions, climate and environmental changes, and low seed germination and plant growth efficiency.
Chitosan is used as a seed initiator, plant growth regulator and resistance inducer, and seedlings and seedlings are treated by soaking seeds and spraying solutions to improve seed germination rate, promote plant growth and enhance stress resistance.
Chitosan significantly improves seed germination rate, promotes plant growth, enhances plant disease resistance, increases plant fresh and dry weight, inhibits disease, enhances antioxidant enzyme activity, and improves stress resistance.
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Abstract
Description
Technical Field
[0001] The present invention belongs to the technical field of agriculture and relates to the application of natural polysaccharides in the preparation of seed priming agents, plant growth regulators and / or resistance inducers. Background Art
[0002] In the production process of watermelon, multiple challenges are often encountered, which are directly related to the decline in yield and quality. The primary problem lies in the soil conditions, and factors such as its quality, fertility and pH value may all pose obstacles to the growth of watermelon. Subsequently, the changes in the climate environment, such as unsuitable temperature and moisture conditions, also become factors restricting the healthy growth of watermelon. In addition, the frequent occurrence of pests and diseases, such as the invasion of Fusarium wilt, anthracnose and pests such as aphids and spider mites, seriously threatens the yield and marketability of watermelon. Given that most of these factors are difficult to control artificially, the current relatively economical and effective countermeasure is to cultivate strong seedlings to improve their survival rate, thereby laying a solid foundation for the high yield and high quality of watermelon.
[0003] Seed germination and plant growth are key links in agricultural production, and their quality and efficiency directly affect the yield and quality of crops. In order to improve the germination rate and growth speed of seeds, scientific research personnel have continuously explored various seed treatment technologies. Summary of the Invention
[0004] To solve the above technical problems, the purpose of the present invention is to provide the application of natural polysaccharides in the preparation of seed priming agents, plant growth regulators and / or resistance inducers.
[0005] The present invention relates to the application of chitosan in the preparation of seed priming agents, plant growth regulators and / or resistance inducers.
[0006] In the present invention, chitosan (English name chitosan) is a natural polysaccharide, a natural, non-toxic and low-cost biopolymer material, in the form of a white powder, odorless and tasteless, with a relatively high molecular weight and a linear structure without branches.
[0007] In the above application, the seed priming agent prepared from chitosan is a priming agent for improving the seed germination rate; the improvement of the seed germination rate is reflected in promoting seed germination and / or improving seed vigor; The plant growth regulator prepared from chitosan is a regulator for promoting plant growth; the promotion of plant growth is reflected in increasing the fresh weight and / or dry weight of the plant; The resistance inducer prepared from chitosan is an inducer for improving plant stress resistance; The improvement of plant stress resistance is reflected in improving plant disease resistance.
[0008] The present invention also provides a seed priming agent, a plant growth regulator and / or a resistance inducer, the active ingredient of which is the chitosan described above.
[0009] In the above-mentioned seed priming agent, plant growth regulator and / or resistance inducer, the priming agent, plant growth regulator or resistance inducer is prepared by dissolving the chitosan in an aqueous solution of an acid. The acid is selected from at least one of acetic acid, carbonic acid, citric acid, hydrosulfuric acid, phosphoric acid and hydrochloric acid.
[0010] In the above-mentioned seed priming agent, plant growth regulator and / or resistance inducer, the volume percentage concentration of the aqueous solution of the acid can be 0.5 - 1.5%. The pH of the priming agent can be 5.5 - 6.5. In the seed priming agent, plant growth regulator or resistance inducer, the concentration of the chitosan can be 600 ± 50 mg / L, specifically 600 mg / L. Generally, the concentration of the mother liquor of the chitosan is 5000 ± 50 mg / L, and it is diluted to 600 ± 50 mg / L for use.
[0011] The present invention also provides a method for improving the seed germination rate, which includes the following steps: during the seed priming treatment, soaking the seeds with the above-mentioned seed priming agent, drying back, and germinating to improve the seed germination rate.
[0012] In the present invention, the seeds can specifically be watermelon seeds.
[0013] In the above-mentioned method, the soaking time is 8 - 13 hours, specifically 12 hours, and the time for the seeds to dry back after soaking is 20 - 36 hours, specifically 24 hours.
[0014] The present invention also provides a method for promoting plant growth, which includes the following steps: during the seed priming treatment, soaking the seeds with the above-mentioned seed priming agent, drying back, germinating and then sowing; during the plant seedling stage, spraying the above-mentioned plant growth regulator to promote the growth of seedlings.
[0015] In the above-mentioned method, the plant seedling stage is the two-leaf and one-heart stage.
[0016] In the present invention, the seeds can specifically be watermelon seeds.
[0017] The present invention further provides a method for improving plant stress resistance, which includes the following steps: during the seed priming treatment, soaking the seeds with the above-mentioned seed priming agent, drying back, germinating and then sowing; during the plant seedling stage, spraying the above-mentioned plant growth regulator, and then transplanting the seedlings to a field with continuous cropping for many years, and spraying and treating with the above-mentioned resistance inducer to improve plant stress resistance.
[0018] In the above method, the seedling stage of the plant can be the two-leaf and one-heart stage; The period for transplanting the seedlings can be the four-leaf stage of the seedlings.
[0019] In the present invention, the seeds can specifically be watermelon seeds.
[0020] In the above method, for the spraying treatment after transplanting the four-leaf stage seedlings to the field with continuous cropping for many years, the roots and their leaves are evenly sprayed, once every 7 days, for a total of 4 times.
[0021] The present invention has the following beneficial effects: 1. Chitosan can promote seed germination, improve seed vigor, and promote early seed germination.
[0022] 2. Chitosan can promote plant growth and improve growth indexes such as plant fresh weight and dry weight.
[0023] 3. Chitosan can inhibit the growth of watermelon fusarium wilt pathogen, and improve the expression level of disease-resistant genes in watermelon, improving the stress resistance of watermelon.
[0024] 4. Chitosan can improve the antioxidant enzyme activity in watermelon plants, improving the stress resistance of watermelon.
[0025] 5. Chitosan can improve the growth potential of watermelon in the field with continuous cropping for many years, and prevent the occurrence of diseases and / or reduce the harm caused by diseases. Description of the Drawings
[0026] Figure 1 For Example 1, the chitosan priming treatment has a relatively high germination rate for watermelon seeds.
[0027] Figure 2 For Example 2, the chitosan priming treatment has a relatively high germination rate for triploid watermelon seeds.
[0028] Figure 3 For Example 3, the chitosan priming treatment has a relatively high germination rate for corn seeds.
[0029] Figure 4 For Example 4, the chitosan priming treatment has a relatively high germination rate for tomato seeds.
[0030] Figure 5 For Example 5, the chitosan priming treatment has a relatively high germination rate for cotton seeds.
[0031] Figure 6 For Example 6, the growth situation of watermelon seedlings treated with chitosan priming is evaluated.
[0032] Figure 7 For Example 7, the growth situation of corn seedlings treated with chitosan priming is evaluated.
[0033] Figure 8 Evaluation of the growth of tomato seedlings treated with chitosan in Example 8.
[0034] Figure 9 Evaluation of the growth of cotton seedlings treated with chitosan in Example 9.
[0035] Figure 10 Inhibitory effect of chitosan on Fusarium oxysporum f. sp. niveum in Example 10.
[0036] Figure 11 Effect of chitosan on disease-resistant genes in watermelon plants in Example 11.
[0037] Figure 12 Effect of chitosan on the activity of antioxidant enzymes in watermelon plants in Example 12.
[0038] Figure 13 Application effect of chitosan in the field in Example 13. Specific implementation mode
[0039] The experimental methods used in the following examples are all conventional methods unless otherwise specified.
[0040] The materials, reagents, etc. used in the following examples can be obtained from commercial sources unless otherwise specified.
[0041] Example 1 As a preparation method of a seed priming agent (also known as chitosan solution), it includes the following steps: Step S1: Weigh 1 g of chitosan, mix chitosan with an acetic acid aqueous solution with a volume concentration of 1%, add distilled water to make up the volume, and obtain a chitosan stock solution of system A with a pH of 5.5 - 6.5 and a stock solution concentration of 5 g / L.
[0042] Step S2: Dilute the stock solution of system A to a chitosan treatment concentration of 600 mg / L, which is the chitosan solution.
[0043] This example provides an application experiment using chitosan solution to improve the germination rate of watermelon seeds. The specific operation steps are as follows: Soak watermelon seeds of the Huaxin variety in water and 600 mg / L chitosan solution respectively for 12 h, and then dry them back at room temperature (25 °C) for 24 h. Place the dried-back watermelon seeds evenly on a culture dish lined with germination paper, with 50 seeds in each culture dish, set three replicates, place them in an incubator at 30 °C, count the number of germinated seeds, and calculate the germination rate. The untreated with chitosan is used as the control group (denoted as CK). The experimental results are as Figure 1 shown, from Figure 1As can be seen from the results, after the chitosan initiation treatment, the germination rate increased by approximately 9%.
[0044] Example 2 The following is the application experiment of using the chitosan solution prepared in Example 1 of the present invention as a seed initiator to improve the germination rate of triploid watermelon seeds. The specific operation steps are as follows: The triploid watermelon seeds were soaked in water and a 600 mg / L chitosan solution respectively for 12 h, and then dried back at room temperature for 24 h. The dried-back triploid watermelon seeds were evenly placed in a petri dish lined with germination paper, with 50 seeds in each petri dish, and three replicates were set. They were placed in an incubator at 30 °C, and the number of germinated seeds was counted to calculate the germination rate. The untreated seeds without chitosan were used as the control group (denoted as CK). The experimental results are as Figure 2 shown. From Figure 2 As can be seen from the results, after the chitosan initiation treatment, the germination rate increased by approximately 15%.
[0045] Example 3 The following is the application experiment of using the chitosan solution prepared in Example 1 of the present invention as a seed initiator to improve the germination rate of corn seeds. The specific operation steps are as follows: The corn seeds of the Qianfeng 339 variety were soaked in water and a 600 mg / L chitosan solution respectively for 12 h, and then dried back at room temperature for 24 h. The dried-back corn seeds were evenly placed in a petri dish lined with germination paper, with 50 seeds in each petri dish, and three replicates were set. They were placed in an incubator at 30 °C, and the number of germinated seeds was counted to calculate the germination rate. The untreated seeds without chitosan were used as the control group (denoted as CK). The experimental results are as Figure 3 shown. From Figure 3 As can be seen from the results, after the chitosan initiation treatment, the germination rate increased by approximately 11.5%.
[0046] Example 4 The following is the application experiment of using the chitosan solution prepared in Example 1 of the present invention as a seed initiator to improve the germination rate of tomato seeds. The specific operation steps are as follows: The tomato seeds of the Huangxing No. 6 variety were soaked in water and a 600 mg / L chitosan solution respectively for 12 h, and then dried back at room temperature for 24 h. The dried-back tomato seeds were evenly placed in a petri dish lined with germination paper, with 50 seeds in each petri dish, and three replicates were set. They were placed in an incubator at 30 °C, and the number of germinated seeds was counted to calculate the germination rate. The untreated seeds without chitosan were used as the control group (denoted as CK). The experimental results are as Figure 4 shown. From Figure 4 As can be seen from the results, after the chitosan initiation treatment, the germination rate increased by approximately 20%.
[0047] Example 5 An application experiment on using the chitosan solution prepared in Example 1 of the present invention as a seed initiator to improve the germination rate of cotton seeds was carried out. The specific operation steps are as follows: Cotton seeds of variety Shen G5 were soaked in clean water and a chitosan solution with a concentration of 600 mg / L respectively for 12 h, and then dried back at room temperature for 24 h. The dried-back cotton seeds were evenly placed in a petri dish lined with germination paper, with 50 seeds in each petri dish, and three replicates were set. They were placed in an incubator at 30 °C, and the number of germinated seeds was counted to calculate the germination rate. The treatment without chitosan was used as the control group (denoted as CK). The experimental results are as Figure 5 shown. From Figure 5 the results, it can be seen that after the chitosan priming treatment, the germination rate increased by about 8%.
[0048] Example 6 An application experiment on using the chitosan solution prepared in Example 1 of the present invention as a plant growth regulator to promote the growth of watermelon seedlings was carried out. The specific operation steps are as follows: Watermelon seeds of variety Huaxin were soaked in clean water and a chitosan solution with a concentration of 600 mg / L respectively for 12 h, and then dried back at room temperature for 24 h. The dried-back watermelon seeds were sown in a plug tray filled with substrate, with three replicates set, and were placed in a greenhouse for cultivation until the two-leaf and one-heart stage. Then, a 600 mg / L chitosan solution was sprayed once, and the plant growth indexes were measured 7 d later. The treatment without chitosan was used as the control group (denoted as CK). The experimental results are as Figure 6 shown. From Figure 6 the results, it can be seen that after the chitosan priming treatment, the plant height, root length, fresh weight, and dry weight all increased significantly. Compared with the control group CK without chitosan treatment, the plant height, root length, fresh weight, and dry weight increased by about 1.3 times, 1.5 times, 2.3 times, and 1.3 times respectively.
[0049] Example 7 An application experiment on using the chitosan solution prepared in Example 1 of the present invention as a plant growth regulator to promote the growth of corn seedlings was carried out. The specific operation steps are as follows: Corn seeds of variety Qianfeng 339 were soaked in clean water and a chitosan solution with a concentration of 600 mg / L respectively for 12 h, and then dried back at room temperature for 24 h. The dried-back corn seeds were sown in a plug tray filled with substrate, with three replicates set, and were placed in a greenhouse for cultivation until the two-leaf and one-heart stage. Then, a 600 mg / L chitosan solution was sprayed once, and the plant growth indexes were measured 7 d later. The treatment without chitosan was used as the control group (denoted as CK). The experimental results are as Figure 7 shown. From Figure 7It can be seen from the results that after the treatment with chitosan initiation, the plant height, root length, fresh weight, and dry weight all increased significantly. Compared with the control group CK without chitosan treatment, the plant height, root length, fresh weight, and dry weight increased by about 1.2 times, 1.3 times, 1.5 times, and 1.1 times respectively.
[0050] Example 8 The application experiment of using the chitosan solution prepared in Example 1 of the present invention as a plant growth regulator to promote the growth of tomato seedlings is as follows: Tomato seeds of the variety Huangxing No. 6 were soaked in water and a 600 mg / L chitosan solution respectively for 12 h, and then dried back at room temperature for 24 h. The dried-back tomato seeds were sown in plug trays filled with substrate, with three replicates, placed in a greenhouse and cultured until the two-leaf and one-heart stage, and sprayed with a 600 mg / L chitosan solution once. The plant growth indexes were measured 7 d later. The group without chitosan treatment was used as the control group (denoted as CK), and the experimental results are as Figure 8 shown. From Figure 8 It can be seen from the results that after the treatment with chitosan initiation, the plant height, root length, fresh weight, and dry weight all increased significantly. Compared with the control group CK without chitosan treatment, the plant height, root length, fresh weight, and dry weight increased by about 1.4 times, 3 times, 1.8 times, and 2.2 times respectively.
[0051] Example 9 The application experiment of using the chitosan solution prepared in Example 1 of the present invention as a plant growth regulator to promote the growth of cotton seedlings is as follows: Cotton seeds of the variety Shen G were soaked in water and a 600 mg / L chitosan solution respectively for 12 h, and then dried back at room temperature for 24 h. The dried-back cotton seeds were sown in plug trays filled with substrate, with three replicates, placed in a greenhouse and cultured until the two-leaf and one-heart stage, and sprayed with a 600 mg / L chitosan solution once. The plant growth indexes were measured 7 d later. The group without chitosan treatment was used as the control group (denoted as CK), and the experimental results are as Figure 9 shown. From Figure 9 It can be seen from the results that after the treatment with chitosan initiation, the plant height, root length, fresh weight, and dry weight all increased significantly. Compared with the control group CK without chitosan treatment, the plant height, root length, fresh weight, and dry weight increased by about 1.2 times, 2 times, 1.4 times, and 1.6 times respectively.
[0052] Example 10 The application experiment of using the chitosan solution prepared in Example 1 of the present invention as a resistance inducer to inhibit the growth of watermelon fusarium wilt pathogen is as follows: The chitosan mother liquor prepared in Example 1 was added with an appropriate amount of PDA medium (added with antibiotic AMP+) in a laminar flow hood, poured into petri dishes, and left standing for 30 min to prepare PDA solid media containing different concentrations of chitosan: 0, 200, 400, 600, 800, 1000 mg / L. The treatment with 0 mg / L was used as the control treatment. A puncher with a diameter of 8 mm was used to transfer the fungal disks into the above-mentioned culture media with different treatments. Starting from Figure 10 As can be seen, chitosan has a significant inhibitory effect on Fusarium oxysporum f. sp. niveum starting from 200 mg / L.
[0053] Example 11 An application experiment of using the chitosan solution prepared in Example 1 of the present invention as a resistance inducer to regulate disease-resistant genes in watermelon plants was carried out. The specific operation steps are as follows: RNA was extracted from the samples treated with chitosan and untreated samples, and the PCR reaction system was prepared using the Novizan UniPeak U ＋ One Step RT-qPCR SYBR Green Kit kit. The PCR reaction system was placed in a real-time fluorescence quantitative PCR instrument, and the PCR amplification reaction was carried out according to the set temperature and time. According to the change of fluorescence signal, the amplification curve was drawn, and the expression level of the target gene was calculated. Starting from Figure 11 The results showed that NPR1, PR1a, PP2C, WRKY30, WRKY46, and PAD increased by about 4.6 times, 2.2 times, 11.2 times, 1.6 times, 1.2 times, and 2.3 times respectively.
[0054] Example 12 An application experiment of using the chitosan solution prepared in Example 1 of the present invention as a resistance inducer to regulate the activities of antioxidant enzymes in watermelon plants was carried out. The specific operation steps are as follows: The activities of SOD and CAT enzymes were measured in the samples treated with chitosan and untreated samples. The specific steps are detailed in the enzyme activity assay kit. The experimental results are as Figure 12 shown. Starting from Figure 12 As can be seen from the results, after treatment with chitosan, the activities of SOD and CAT enzymes were significantly increased. Compared with the control group CK without chitosan treatment, the activities of SOD and CAT enzymes increased by about 6.4 times and 18 times respectively.
[0055] Example 13 An application experiment of using the chitosan solution prepared in Example 1 of the present invention as a resistance inducer to regulate the growth and stress resistance of field watermelon was carried out. The specific operation steps are as follows: Before seed germination, soak the seeds with the above-mentioned chitosan solution to promote seed germination. At the two-leaf and one-heart stage, spray the chitosan solution to promote seedling growth. At the four-leaf stage, transplant the seedlings to the field with continuous cropping for years and spray the chitosan solution. The experimental results are as Figure 13 shown. It can be seen from the field phenotypes that chitosan can significantly promote the growth of watermelon seedlings and effectively prevent the occurrence of diseases and / or reduce the harm caused by diseases.
Claims
1. Application of chitosan in preparing seed initiator, plant growth regulator and / or resistance inducer.
2. The application according to claim 1, wherein The seed initiator prepared from the chitosan is an initiator for increasing seed germination rate; the improvement of seed germination rate is reflected in promoting seed germination and / or enhancing seed vigor; The plant growth regulator prepared from the chitosan is a regulator for promoting plant growth; the promotion of plant growth is reflected in increasing plant fresh weight and / or dry weight; The resistance inducer prepared from the chitosan is an inducer for enhancing plant disease resistance; The improvement of plant stress resistance is reflected in inhibiting the growth of Fusarium oxysporum, increasing the expression level of disease-resistant genes and / or enhancing the activity of antioxidant enzymes in plants.
3. A seed initiator, plant growth regulator, and / or resistance inducer, characterized in that, Its active ingredient is the chitosan described in claim 1 or 2.
4. The seed initiator, plant growth regulator and / or resistance inducer according to claim 3, characterized in that, The said initiator, plant growth regulator or resistance inducer is prepared by dissolving the said chitosan in an aqueous solution of an acid; The said acid is selected from at least one of acetic acid, carbonic acid, citric acid, hydrosulfuric acid, phosphoric acid and hydrochloric acid.
5. The seed initiator, plant growth regulator and / or resistance inducer according to claim 3 or 4, characterized in that, The volume percentage concentration of the aqueous solution of the said acid is 0.5 - 1.5%; The pH of the said initiator is 5.5 - 6.5; The concentration of the chitosan in the said seed initiator, plant growth regulator or resistance inducer is 600 ± 50 mg / L.
6. A method for increasing seed germination rate, comprising the following steps: when performing seed priming treatment, soaking the seeds with the seed initiator described in any one of claims 3 - 5, drying back, and germinating to increase the seed germination rate.
7. A method for promoting plant growth, comprising the following steps: when performing seed priming treatment, soaking the seeds with the seed initiator described in any one of claims 3 - 5, drying back, germinating and then sowing; during the plant seedling stage, spraying the plant growth regulator described in any one of claims 3 - 5 to promote seedling growth.
8. The method according to claim 7, wherein The said plant seedling stage is the two-leaf and one-heart stage.
9. A method for enhancing plant stress resistance, comprising the following steps: when performing seed priming treatment, soaking the seeds with the seed initiator described in any one of claims 3 - 5, drying back, germinating and then sowing; during the plant seedling stage, spraying the plant growth regulator described in any one of claims 3 - 5, and then transplanting the seedlings to a field with continuous cropping for years, and spraying and treating with the resistance inducer described in any one of claims 3 - 5 to enhance plant stress resistance.
10. The method according to claim 9, characterized in that, The said plant seedling stage is the two-leaf and one-heart stage; The period of transplanting the seedlings is the four-leaf stage of the seedlings.