A complex composition of palmitic acid and berberine and use thereof

Abstract

The present disclosure relates to a composition comprising palmitic acid and berberine complex, which can effectively improve the drought resistance of crops and promote the growth and development of crops under drought stress. The present disclosure also provides the use of the composition and a method for improving the growth and development of crops under drought stress.

Description

Technical Field

[0001] This invention relates to the fields of plant stress physiological regulation and agricultural chemistry, specifically to an exogenous compound composition consisting of fatty acid compounds and amine compounds and its application in improving crop drought resistance and promoting crop growth. Background Technology

[0002] Existing drought-resistant agents are mostly chemically synthesized substances, and often work well for single crops, but their environmental compatibility remains a concern. Furthermore, the need to apply different drought-resistant agents to different crops increases the complexity of management in actual agricultural production. Therefore, there is an urgent need to develop a composition that is relatively environmentally friendly, suitable for multiple crops, and can improve crop drought resistance. Summary of the Invention

[0003] To address the problems existing in the prior art, this disclosure provides a composition for improving crop drought resistance and / or promoting crop growth and development, the composition comprising palmitic acid and berberine.

[0004] In some embodiments, the main components of the composition consist of palmitic acid and berberine.

[0005] In some embodiments, the mass ratio of palmitic acid to berberine is (6-10):1, preferably 7.6:1.

[0006] In some embodiments, the concentration of palmitic acid is 0.20-1.00 mmol / L, and the concentration of berberine is 0.05-0.20 mmol / L.

[0007] In some embodiments, the concentration of palmitic acid is 1.00 mmol / L and the concentration of berberine is 0.10 mmol / L.

[0008] In some embodiments, the composition can be used as the main active ingredient in fertilizers or plant growth regulators.

[0009] In some embodiments, the composition is applied by foliar spraying during the seedling stage.

[0010] In some embodiments, the crop is flax, corn, soybean, or sorghum.

[0011] This disclosure also provides a fertilizer for improving crop drought resistance and / or promoting crop growth and development, wherein the active ingredients of the fertilizer include palmitic acid and berberine.

[0012] In some embodiments, the mass ratio of palmitic acid to berberine is (6-10):1, preferably 7.6:1.

[0013] In some embodiments, the concentration of palmitic acid is 0.20-1.00 mmol / L, and the concentration of berberine is 0.05-0.20 mmol / L.

[0014] In some embodiments, the concentration of palmitic acid is 1.00 mmol / L and the concentration of berberine is 0.10 mmol / L.

[0015] In some embodiments, the fertilizer is a foliar fertilizer.

[0016] In some embodiments, the fertilizer also includes nitrogen, phosphorus, potassium and / or trace elements required for plant growth.

[0017] In some embodiments, the nitrogen can be ammonium nitrate, ammonium bicarbonate, etc., the phosphorus can be superphosphate, monoammonium phosphate, etc., the potassium can be potassium chloride, potassium sulfate, potassium nitrate, etc., and the trace elements can be zinc sulfate, ferrous sulfate, etc.

[0018] In some embodiments, the fertilizer also includes one or more drought-resistant active substances such as proline, betaine, seaweed extract, amino acids, humic acid, or fulvic acid.

[0019] In some embodiments, the crop is flax, corn, soybean, or sorghum.

[0020] In addition, the uses of the compositions or fertilizers described in this disclosure in improving crop drought resistance and / or promoting crop growth and development are also provided.

[0021] In some embodiments, the crop is flax, corn, soybean, or sorghum.

[0022] This disclosure also includes a method for improving crop drought resistance and / or promoting crop growth and development, characterized by applying the composition or fertilizer described in this disclosure to the crop.

[0023] In some embodiments, the compositions or fertilizers described herein are applied by foliar spraying when the crop has just entered the seedling stage.

[0024] In some embodiments, the composition or fertilizer is applied by spraying once every 3 days for 10 consecutive times, with a single spray dose of 0.05–0.20 L / m² leaf area.

[0025] In some embodiments, the composition or fertilizer is sprayed at least 10 times consecutively.

[0026] In some implementations, the single application dose is 0.10 L / m² leaf area.

[0027] In some implementations, the spray is applied to both sides of the leaf to ensure even wetting without significant dripping.

[0028] In some embodiments, the crop is flax, corn, soybean, or sorghum.

[0029] In some embodiments, the composition or fertilizer can be applied during the crop seedling stage.

[0030] The beneficial effects of this disclosure are as follows: This disclosure creatively discovers that the plant metabolites palmitic acid and berberine can effectively improve crop drought resistance and promote crop growth and development. Through the combined application of palmitic acid and berberine, under drought conditions, it can significantly improve crop membrane stability and cellular metabolic homeostasis, enhance crop tolerance to drought stress, and significantly increase crop height and dry weight, thereby improving crop growth performance and increasing yield potential. It is applicable to various crops such as soybeans, corn, sorghum, and flax. Furthermore, since both palmitic acid and berberine are plant metabolites, their application will not cause environmental pollution, making it of great value for widespread application. Attached Figure Description

[0031] Figure 1 The images show maize phenotypic figures. The left image represents the moisture control group, and the right image represents the treatment group treated with palmitic acid and berberine.

[0032] Figure 2 The images show soybean phenotypes. The left image represents the moisture control group, and the right image represents the treatment group treated with palmitic acid and berberine.

[0033] Figure 3 The images show phenotypic figures of sorghum. The left image shows the moisture control group, and the right image shows the treatment group treated with palmitic acid and berberine.

[0034] Figure 4 The images show flax phenotypic figures. The left image represents the moisture control group, and the right image represents the treatment group treated with palmitic acid and berberine. Detailed Implementation

[0035] The technical solutions in the embodiments of this disclosure will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of this disclosure, and not all of them. Based on the embodiments in this disclosure, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of this disclosure. In addition, the technical solutions of various embodiments can be combined with each other, but this must be based on the ability of those of ordinary skill in the art to implement them. When the combination of technical solutions is contradictory or cannot be implemented, it should be considered that such combination of technical solutions does not exist and is not within the scope of protection claimed by this disclosure.

[0036] Unless otherwise specified, all reagents used in this disclosure are commercially available reagents; unless otherwise specified, all techniques and experimental methods used in this disclosure are conventional techniques and methods in the field.

[0037] Previous field drought studies have found that the plant metabolites palmitic acid and berberine are closely related to the drought resistance and growth process of flax. To verify whether palmitic acid and berberine can improve the drought resistance of various crops, the following experiments were conducted.

[0038] Palmitic acid and berberine were both purchased from Nanjing Huiya Biotechnology Co., Ltd. Palmitic acid product code: P815432-1g; Berberine product code: B878903-5mg.

[0039] The preparation method of the palmitic acid and berberine mixture in the following examples is as follows: palmitic acid is dissolved in a small amount of ethanol or dimethyl sulfoxide to form a mother liquor, and berberine is dissolved in water to form an aqueous solution. 0.01%–0.1% Tween-20 is added to water, stirred, and then the above two solutions are added and the volume is adjusted to obtain the spraying mixture, wherein the concentration of palmitic acid is 1.00 mmol / L and the concentration of berberine is 0.10 mmol / L.

[0040] Example 1: Changes in maize plant height and dry weight after spraying with a combination of palmitic acid and berberine.

[0041] Corn variety: Selected corn ( Zea mays L.) self-crossing line B73 was used as the experimental material.

[0042] Seedling cultivation method: After surface sterilization, corn seeds were sown in plug trays (nutrient soil: vermiculite: perlite = 2:1:1, v / v / v) and cultivated under greenhouse conditions. Seedlings grew for 18–21 days (approximately 3 weeks) under normal watering conditions, during which the soil was kept moist. When the seedlings reached the three-leaf stage (V3 stage), 6 uniformly growing seedlings were retained per pot for subsequent metabolite spraying and drought stress experiments. During cultivation, normal watering was provided, the substrate was kept moist, and no stress treatments were applied.

[0043] Pretreatment method for seedling drought stress: Water each pot until the soil is saturated, and allow excess water to drain naturally to ensure that the initial soil moisture content of each pot is consistent. Then, begin the drought treatment.

[0044] Drought stress conditions: In the greenhouse pot experiment, a single drought treatment level was set up. The greenhouse day / night temperature was controlled at 28 / 20 ℃ (±2 ℃). During the drought treatment, the soil moisture content of the potted plants was maintained at 45–50% of field capacity by controlling the watering frequency. Foliar spraying was carried out simultaneously with the application of drought stress. Foliar spraying was carried out once every 3 days from the start of the drought treatment, for a total of 10 consecutive sprays. The spray solution was a mixed solution of palmitic acid and berberine, with the application concentration of palmitic acid at 1.00 mmol / L and the application concentration of berberine at 0.10 mmol / L. The water control group was sprayed with the same dose of water without palmitic acid and berberine under the same drought conditions. The palmitic acid control group was sprayed with the same dose of aqueous solution containing 1.00 mmol / L palmitic acid without berberine under the same drought conditions.

[0045] After treatment, the plant height and dry weight of the maize seedlings were measured. Plant height was measured using a ruler, starting from the surface of the substrate (the junction of the main stem and the substrate), along the main stem to the base of the ligule of the uppermost fully expanded leaf, measured in cm. After harvesting the seedlings, the plants were removed from the substrate, and the substrate adhering to the root surface was removed. If necessary, the roots were quickly rinsed with clean water and the surface moisture was absorbed. The above-ground and underground parts were combined as a whole plant sample and dried in a 65℃ oven to constant weight. The dry weight of the whole plant was recorded.

[0046] The results showed that before drought treatment, there were no significant differences in plant height and other growth indicators between the treatment group and the control group, indicating that the seedlings in the three groups had the same growth baseline at the beginning of the experiment. The results after drought treatment are shown in Table 1. Compared with the simple drought treatment, the average plant height of the maize in the treatment group reached 78.47 cm, an increase of 11.41% compared with the water control group's 70.43 cm, and an increase of 5.10% compared with the palmitic acid control group. Meanwhile, the increase in plant dry weight was even more significant: the average dry weight of the treatment group was 2.36 g, an increase of approximately 29.67% compared with the water control group's 1.82 g, and an increase of approximately 10.28% compared with the palmitic acid control group. Both the plant height and plant dry weight of the treatment group were significantly different from those of the control groups.

[0047] Table 1. Maize plant height and dry weight

[0048]

[0049] In addition, the phenotypes of the treatment group and the water control group were observed. Phenotypic analysis showed that, under drought conditions, the plant height in the treatment group sprayed with a mixed solution of palmitic acid and berberine was significantly higher than that in the control group, and the leaf spread and overall growth were significantly better. The control group plants generally exhibited slow growth and a higher degree of leaf wilting. Figure 1 The left side shows the moisture control group. Figure 1 (The right side represents the treatment group). These results demonstrate that spraying the aforementioned mixed solution can significantly promote maize growth and biomass accumulation under drought conditions, and helps enhance the drought resistance of seedlings.

[0050] Example 2: Changes in soybean plant height and dry weight after spraying with a combination of palmitic acid and berberine.

[0051] Soybean variety: Heihe 43.

[0052] Seedling cultivation method: Soybean seeds were surface-sterilized and sown in plug trays (nutrient soil: vermiculite: perlite = 2:1:1, v / v / v) and cultivated under greenhouse conditions. During cultivation, normal watering was maintained to keep the soil moist. Seedlings were cultivated for 12–14 days (approximately 2 weeks), until they reached the stage of full expansion of the first compound leaf (V1 stage). Six uniformly growing seedlings were retained from each pot for subsequent experiments. Normal watering and substrate moisture were maintained during cultivation, and no stress treatments were applied.

[0053] Pretreatment method for seedling drought stress: Water each pot until the soil is saturated, and allow excess water to drain naturally to ensure that the initial soil moisture content of each pot is consistent before starting the drought treatment.

[0054] Drought stress conditions: In the greenhouse pot experiment, a single drought treatment level was set up. The greenhouse day / night temperature was controlled at 28 / 20 ℃ (±2 ℃). During the drought treatment, the soil moisture content of the potted plants was maintained at 45–50% of field capacity by controlling the watering frequency. Foliar spraying was carried out simultaneously with the application of drought stress. Starting from the day the drought treatment began, the plants in the treatment group were sprayed with the aforementioned metabolite mixture solution once every 3 days for 10 consecutive times. The spray solution was a mixture of palmitic acid and berberine, with the application concentration of palmitic acid at 1.00 mmol / L and the application concentration of berberine at 0.10 mmol / L. The water control group was sprayed with the same dose of water without palmitic acid and berberine under the same drought conditions. The palmitic acid control group was sprayed with the same dose of aqueous solution containing 1.00 mmol / L palmitic acid without berberine under the same drought conditions.

[0055] After treatment, the plant height and dry weight of soybean seedlings were measured. Plant height was measured using a ruler, starting from the surface of the soybean seedling substrate (the junction of the main stem and substrate), and extending along the main stem to the apex of the main stem, measured in cm. After harvesting the soybean seedlings, the plants were removed from the substrate, and the substrate adhering to the root surface was removed. If necessary, the roots were quickly rinsed with clean water and the surface moisture was absorbed. The above-ground and underground parts were combined as a whole plant sample and dried in a 65℃ oven until constant weight. The dry weight of the whole plant was recorded.

[0056] The results showed that before drought treatment, there were no significant differences in plant height and other growth indicators between the treatment group and the control groups, indicating that the seedlings in the three groups had the same growth baseline at the beginning of the experiment. As shown in Table 2, compared with the simple drought treatment, the average plant height of soybeans in the treatment group reached 64.64 cm, which was 9.50% higher than the 59.03 cm in the water control group and 4.41% higher than the palmitic acid control group. The average plant dry weight was 0.61 g, which was 35.56% higher than the 0.45 g in the water control group and 15.09% higher than the palmitic acid control group. The plant height and plant dry weight of the treatment group were significantly different from those of the control groups.

[0057] Table 2 Soybean plant height and plant dry weight

[0058]

[0059] In addition, the phenotypes of the treatment group and the water control group were observed. Phenotypic analysis showed that, under drought conditions, the plant height in the treatment group sprayed with a mixed solution of palmitic acid and berberine was significantly higher than that in the control group, and the leaf spread and overall growth were significantly better. The control group plants generally exhibited slow growth and a higher degree of leaf wilting. Figure 2 The left side shows the moisture control group. Figure 2 (The right side represents the treatment group). These results demonstrate that the combined application of the aforementioned mixed solution of palmitic acid and berberine can significantly promote the height growth and dry matter accumulation of soybean plants, thereby enhancing their growth potential and adaptability under drought conditions.

[0060] Example 3: Changes in sorghum plant height and dry weight after spraying with a combination of palmitic acid and berberine.

[0061] Sorghum variety: Jixiangliang No. 1.

[0062] Seedling cultivation method: Sorghum seeds were surface-sterilized and sown in plug trays filled with nutrient soil (nutrient soil: vermiculite: perlite = 2:1:1, v / v / v). Seedlings were cultivated in a greenhouse for 20–24 days (approximately 3 weeks), until they reached the stage where the third true leaf was fully expanded (three-leaf stage, V3 stage). Six uniformly growing seedlings were retained from each pot for subsequent experimental treatments. During the cultivation period, normal watering was provided to keep the substrate moist, and no stress treatments were applied.

[0063] Pretreatment method for seedling drought stress: Water each pot until the soil is saturated, and allow excess water to drain naturally to ensure that the initial soil moisture content of each pot is consistent before starting the drought stress treatment.

[0064] Drought stress conditions: In the greenhouse pot experiment, a single drought treatment level was set up. The greenhouse day / night temperature was controlled at 28 / 20 ℃ (±2 ℃). During the drought treatment, the soil moisture content of the potted plants was maintained at 45–50% of field capacity by controlling the watering frequency. Foliar spraying was carried out simultaneously with the application of drought stress. Starting from the day the drought treatment began, the plants in the treatment group were sprayed with the aforementioned metabolite mixture solution once every 3 days for 10 consecutive times. The spray solution was a mixture of palmitic acid and berberine, with the application concentration of palmitic acid at 1.00 mmol / L and the application concentration of berberine at 0.10 mmol / L. The water control group was sprayed with the same dose of water without palmitic acid and berberine under the same drought conditions. The palmitic acid control group was sprayed with the same dose of aqueous solution containing 1.00 mmol / L palmitic acid without berberine under the same drought conditions.

[0065] After treatment, the plant height and dry weight of the sorghum seedlings were measured. Plant height was measured using a ruler, starting from the surface of the substrate (the junction of the main stem and the substrate) and extending along the main stem to the apical meristem, measured in cm. After harvesting the sorghum seedlings, the plants were removed from the substrate, and the substrate adhering to the root surface was removed. If necessary, the roots were quickly rinsed with clean water and the surface moisture was absorbed. The above-ground and underground parts were combined as a whole plant sample and dried in a 65℃ oven to constant weight. The dry weight of the whole plant was recorded.

[0066] The results showed that before drought treatment, there were no significant differences in plant height and other growth indicators between the treatment group and the control groups, indicating that the seedlings in the three groups had the same growth baseline at the beginning of the experiment. As shown in Table 3, after drought treatment, compared with the simple drought treatment, the average plant height of sorghum in the treatment group reached 67.28 cm, which was 9.92% higher than the 61.21 cm in the water control group and 4.44% higher than the palmitic acid control group. The average plant dry weight was 0.43 g, which was 19.44% higher than the 0.36 g in the water control group and 10.26% higher than the palmitic acid control group. The plant height and plant dry weight of the treatment group were significantly different from those of the control groups.

[0067] Table 3 Sorghum plant height and dry weight

[0068]

[0069] In addition, the phenotypes of the treatment group and the water control group were observed. Phenotypic analysis showed that, under drought conditions, the plant height in the treatment group sprayed with a mixed solution of palmitic acid and berberine was significantly higher than that in the control group, and the leaf spread and overall growth were significantly better. The control group plants generally exhibited slow growth and a higher degree of leaf wilting. Figure 3 The left side shows the moisture control group. Figure 3 (The right side shows the treatment group). These results indicate that spraying the mixed solution can significantly improve the growth performance of sorghum under drought conditions, promote plant height increase and dry matter accumulation, and help enhance its adaptability to drought environments.

[0070] Example 4: Changes in flax plant height and dry weight after spraying with a combination of palmitic acid and berberine.

[0071] Flax variety: R43

[0072] Seedling Cultivation Method: Preliminary screening revealed that flax seedlings did not grow well in plug trays; therefore, flax was cultured in pots in subsequent experiments. Flax seeds were surface-sterilized and sown in pots filled with nutrient soil (nutrient soil: vermiculite: perlite = 2:1:1, v / v / v). Seedlings were cultivated in a greenhouse for 18–22 days (approximately 3 weeks), until they reached the stage of full expansion of the first pair of true leaves to the stage of expansion of the third true leaf (BBCH 12–13). Ten uniformly growing seedlings were retained from each pot for subsequent experimental treatments. Normal watering was maintained during cultivation to keep the substrate moist, and no stress treatments were applied.

[0073] Pretreatment method for seedling drought stress: Water each pot until the soil is completely saturated, and allow excess water to drain naturally from the bottom of the pot to ensure that the initial soil moisture content of each pot is consistent.

[0074] Drought stress conditions: In the greenhouse pot experiment, a single drought treatment level was set up. The greenhouse day / night temperature was controlled at 28 / 20 ℃ (±2 ℃). During the drought treatment, the soil moisture content in the 0–20 cm soil layer of flax under field conditions was graded, and the amplification effect of root volume restriction and enhanced evaporation on drought stress perception under greenhouse pot conditions was considered. The water conditions were equivalently adjusted to maintain the soil moisture content in the pot at 40–45% of field capacity by controlling the watering frequency. Foliar spraying was carried out simultaneously with the application of drought stress. Foliar spraying was carried out once every 3 days from the start of the drought treatment, for a total of 10 times. The spray solution was a mixture of palmitic acid and berberine, with the application concentration of palmitic acid at 1.00 mmol / L and the application concentration of berberine at 0.10 mmol / L. The water control group was sprayed with the same dose of water without palmitic acid and berberine under the same drought conditions. The palmitic acid control group consisted of an aqueous solution containing 1.00 mmol / L palmitic acid, which was sprayed under the same drought conditions and did not contain berberine.

[0075] After treatment, the plant height and dry weight of the flax seedlings were measured. Plant height was measured using a ruler, starting from the surface of the substrate (the junction of the main stem and the substrate) and extending along the main stem to the apical meristem, measured in cm. After harvesting the flax seedlings, the plants were removed from the substrate, and the substrate adhering to the root surface was removed. If necessary, the roots were quickly rinsed with clean water and the surface moisture was absorbed. The above-ground and underground parts were combined as a whole plant sample and dried in a 65℃ oven to constant weight. The dry weight of the whole plant was recorded.

[0076] The results showed that before drought treatment, there were no significant differences in plant height and other growth indicators between the treatment group and the control groups, indicating that the seedlings in the three groups had the same growth baseline at the beginning of the experiment. As shown in Table 4, after drought treatment, compared with the simple drought treatment, the average plant height of the treatment group was 34.23 cm, an increase of approximately 15.64% compared to the water control group's 29.60 cm, and an increase of approximately 7.54% compared to the palmitic acid control group. The average plant dry weight reached 0.39 g, an increase of approximately 14.71% compared to the water control group's 0.34 g, and an increase of approximately 8.33% compared to the palmitic acid control group. The plant height and plant dry weight of the treatment group were significantly different from those of the control groups.

[0077] Table 4 Flax plant height and plant dry weight

[0078]

[0079] In addition, the phenotypes of the treatment group and the water control group were observed. Phenotypic analysis showed that, under drought conditions, the plant height in the treatment group sprayed with a mixed solution of palmitic acid and berberine was significantly higher than that in the control group, and the leaf spread and overall growth were significantly better. The control group plants generally exhibited slow growth and a higher degree of leaf wilting. Figure 4 The left side shows the moisture control group. Figure 4 (The right side represents the treatment group). These results indicate that spraying the mixed solution can effectively promote the growth and development of flax under drought stress, improve the plant's biomass accumulation capacity, and thus enhance its adaptability to adverse environments.

[0080] In summary, this disclosure provides a compound composition comprising palmitic acid and berberine. Foliar application of this composition during the seedling stage can significantly enhance the drought resistance of crops under arid conditions and improve plant growth performance. The technical solution has demonstrated stable drought-resistant and growth-promoting effects on crops such as soybeans, corn, sorghum, and flax, significantly increasing plant height and biomass accumulation, thereby contributing to yield formation. This disclosed technical solution has advantages such as simple implementation, wide applicability, and stable effects, making it suitable for agricultural production and stress-resistant cultivation in arid and semi-arid regions, and possesses significant potential for widespread application.

Claims

1. The use of the composition in promoting crop growth and development under drought conditions, characterized in that, The composition comprises palmitic acid and berberine, wherein the concentration of palmitic acid is 1.00 mmol / L and the concentration of berberine is 0.10 mmol / L, and the crop is flax, corn, soybean or sorghum.

2. A method for promoting crop growth and development under drought conditions, characterized in that, The crop is flax, corn, soybean or sorghum, and the palmitic acid is applied to the crop at a concentration of 1.00 mmol / L and the berberine is at a concentration of 0.10 mmol / L.

3. The method as described in claim 2, characterized in that, The composition is applied by foliar spraying when the crop is just entering the seedling stage.

4. The method according to any one of claims 2-3, characterized in that, The composition is applied by spraying once every 3 days for 10 consecutive times, with a single spray dose of 0.05–0.20 L / m² leaf area.

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