Application of borage extract in promoting plant growth and / or regulating plant stress resistance

Abstract

Provided is an application of a borage extract in promoting plant growth and / or regulating plant stress resistance in the technical field of agriculture. At present, the borage extract is only reported to be used in the field of medicine, and not in agriculture. Provided is an application of the borage extract, in particular, the use as a biostimulant for promoting plant growth and enhancing plant stress resistance.

Description

Application of Borage extract in promoting plant growth and / or regulating plant stress resistance Technical Field

[0001] The present invention belongs to the technical field of agriculture, and particularly relates to application of Borage extract in promoting plant growth and / or regulating plant stress resistance. Background Art

[0002] Borage, a member of the Boraginaceae family, is a robust, annual herb. It grows up to 120 cm tall as an annual. The plant is covered in rough, grayish-white hairs. The stem is cylindrical, about 1 1 / 2 feet tall, and has branches. The branches are hollow and rich in water. The dark green, wrinkled leaves are arranged alternately on the stem. They are about 3 inches long and 1 1 / 2 inches wide. The leaves are rigidly and gradually contract into an ellipsoidal shape from bottom to top. The burrs covering the upper and lower leaves and the stem are tipped with tiny rounded tips, giving the outer spherical shape a continuously wavy edge. The fruit contains four dark brown nuts. The leaves are alternate, rough, cucumber-like, and petiolate. The flowers bloom in July in loose, leafy cymes of blue, with long petioles. The nuts are smooth or have papillate projections.

[0003] Borage and / or Borage extracts have certain nutritional value and physiological functions, and are commonly used in the medical field, mainly including: (1) prevention and treatment of cardiovascular diseases; Borage and / or Borage extracts can inhibit angiotensin synthesis, thereby lowering blood pressure, and reducing serum cholesterol and triglyceride levels; (2) treatment of diabetes; Borage and / or Borage extracts have the effect of lowering blood sugar, enhancing the sensitivity of cell membrane receptors to insulin, and protecting the stability of insulin in the blood; (3) treatment of cancer; (4) relief of premenstrual syndrome; (5) treatment of skin diseases; (6) protection of the liver; (7) regulation of immune function, etc. However, it has not been found that Borage extracts are used to promote plant growth.

[0004] Summary of the Invention

[0005] The present invention discovers for the first time that Borage extract not only promotes plant growth but also enhances plant stress resistance, thereby expanding the application scope of Borage extract and providing a new option for regulating plant growth in agriculture.

[0006] The present invention provides the use of Borage extract in promoting plant growth and / or regulating plant stress resistance.

[0007] In the present invention, the Borage extract can promote seed germination and / or the growth of one or more of roots, stems, leaves, or flowers and fruits. For example, it can promote or inhibit root length, stem diameter, plant height, leaf width, leaf length, leaf number, leaf area, biomass, chlorophyll content, yield, and the like.

[0008] Plant stress resistance refers to certain traits that plants possess to resist adverse environments, such as cold resistance, drought resistance, saline-alkali resistance, and high temperature resistance.

[0009] In the present invention, the stress resistance includes but is not limited to saline-alkali resistance, cold resistance, drought resistance, and high temperature resistance.

[0010] In the present invention, the Borage extract is an alcohol extract.

[0011] In the present invention, the alcohol solution is selected from methanol, ethanol and propanol.

[0012] In a specific embodiment of the present invention, the alcohol solution is ethanol or methanol.

[0013] In the present invention, the ethanol concentration or methanol concentration can be selected from 10% to 100%, can be selected from 10% to 20%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, 90% to 100%, and can also be selected from.......30%,.....40%,.....50%,......60%,....70%,....80% and the like.

[0014] Furthermore, the concentration of the ethanol is 40% to 80%.

[0015] In the present invention, the mass ratio of the borage to the ethanol is 1-5:15-25.

[0016] In a specific embodiment of the present invention, the mass ratio of borage to ethanol is 1:20.

[0017] When mass ratio or other value or parameter are expressed with range, preferred range or the range that a series of upper preferred value and lower preferred value limit are expressed, this should be understood as specifically disclosing all ranges formed by any pairing of any range upper limit or preferred value and any range lower limit or preferred value, no matter whether this range is disclosed separately.For example, when disclosing scope "1 to 5", described scope should be interpreted as including scope "1 to 4", "1 to 3", "1 to 2", "1 to 2 and 4 to 5", "1 to 3 and 5" etc.When numerical range is described in this article, unless otherwise stated, otherwise this scope is intended to include its end value and all integers and fractions within this scope.

[0018] In the present invention, the borage extract is used as an active ingredient of the biostimulant and can be used alone or in combination with other products.

[0019] The invention provides a biostimulant which takes borage extract as an active ingredient.

[0020] The term "biostimulant" as used herein includes substances and / or microorganisms that, when applied to plants or the rhizosphere, function to stimulate natural processes to promote and / or inhibit nutrient uptake, increase and / or decrease nutrient conversion efficiency, abiotic tolerance, and crop yield.

[0021] In the present invention, the borage extract is a natural biostimulant.

[0022] The plants described in the present invention include but are not limited to economic crops and food crops, such as tobacco, corn, rice, cucumber, lettuce, wheat, pepper, Chinese cabbage, lettuce, bok choy, tomato, citrus, kiwi, cherry, pear, apple, etc.

[0023] The so-called "economic crops" are of various types, including but not limited to fiber crops (such as cotton, hemp, etc.), oil crops (such as sesame, peanuts, etc.), sugar crops (such as sugarcane, sugar beets, etc.), hobby crops (tobacco), medicinal crops, dye crops, ornamental crops, fruits (such as citrus, kiwi, cherry, pear, apple, etc.) and other economic crops (such as lettuce, pepper, Chinese cabbage, lettuce, Shanghai green, etc.).

[0024] The “food crops” mentioned above include but are not limited to cereal crops (wheat, rice, corn), tuber crops (including sweet potatoes, potatoes, etc.) and legume crops (including soybeans, broad beans, peas, mung beans, etc.).

[0025] In the product of the present invention, the borage extract can be used directly as a single dose. In order to stabilize it and facilitate transportation and storage, it can be prepared into an agricultural product, for example, by adding excipients to prepare a corresponding dosage form. The excipients can be conventional excipients in the art, such as: thickeners, defoamers, dispersants, wetting agents, binders, emulsifiers, stabilizers, solvents, etc.

[0026] On the other hand, the Borage extract of the present invention can be used as a synergist in combination with products such as plant regulators, foliar fertilizers, water-soluble fertilizers, compound fertilizers, and pesticides.

[0027] In the present invention, the dosage form of the agricultural product includes but is not limited to powder, granule, aqueous solution, mother liquor or mother powder.

[0028] In the present invention, when used, the single agent or agricultural product prepared from the borage extract is used to treat seeds, spray on leaves or irrigate roots.

[0029] In the present invention, when the borage extract or its product is prepared into a solution for promoting seed germination or plant growth, the concentration of the borage extract in the solution can be selected according to actual needs.

[0030] For example, it can be selected from 0.01 to 2000 ppm, 0.01 to 1000 ppm, 0.01 to 500 ppm, or 0.01 ppm, ...0.02 ppm, ...0.03 ppm, ...0.1 ppm, ...0.2 ppm, ...0.3 ppm, ...1.0 ppm, ...2.0 ppm, ...3.0 ppm, ...10 ppm, ...20 ppm, ...30 ppm, and the like.

[0031] In some specific embodiments of the present invention, the concentration of the Borage extract is 0.001 ppm to 100 ppm.

[0032] In some specific embodiments of the present invention, the concentration of the Borage extract is 2 to 50 ppm.

[0033] The seeds are treated by mixing a single agent or agricultural product made of borage extract with seeds to coat the seeds with borage extract, or soaking seeds to be germinated with borage extract solution.

[0034] In the growth promotion test of the present invention, the effects of pests and diseases on plant growth have been excluded. Through such an experimental design, the results are sufficient to show that the effect of Borage extract on plant growth promotion is not caused by avoiding pests and diseases.

[0035] The present invention also provides a method for promoting plant growth and / or regulating plant stress resistance by using the borage extract, which comprises applying the borage extract to the plant.

[0036] Beneficial effects of the present invention: The present invention discovers for the first time that borage extract can promote plant growth and enhance its stress resistance, thereby expanding the application scope of borage extract and providing a new option for regulating plant growth in agriculture. DETAILED DESCRIPTION

[0037] The following is a clear and complete description of the technical solution of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0038] Example 1

[0039] Weigh a certain amount of Borage leaf powder, add 20 times the volume of ethanol, the ethanol concentration is 10%, and ultrasonicate at room temperature for 30 minutes. Repeat the above operation twice, combine the filtrates, and concentrate under reduced pressure to a ratio of 1:2 (i.e., 1 g of medicinal material corresponds to 2 g of extract) to obtain Borage extract 1.

[0040] Example 2

[0041] Weigh a certain amount of Borage leaf powder, add 20 times the volume of ethanol, the ethanol concentration is 20%, and ultrasonicate at room temperature for 30 minutes. Repeat the above operation twice, combine the filtrates, and concentrate under reduced pressure to a ratio of 1:2 (i.e., 1 g of medicinal material corresponds to 2 g of extract) to obtain Borage extract 2.

[0042] Example 3

[0043] Weigh a certain amount of Borage leaf powder, add 20 times the volume of ethanol to a concentration of 30%, and sonicate at room temperature for 30 minutes. Repeat the above operation twice, combine the filtrates, and concentrate under reduced pressure to a ratio of 1:2 (i.e., 1 g of medicinal material corresponds to 2 g of extract) to obtain Borage extract 3.

[0044] Example 4

[0045] Weigh a certain amount of Borage leaf powder, add 20 times the volume of ethanol, the ethanol concentration is 40%, ultrasonicate at room temperature for 30 minutes, repeat the above operation twice, combine the filtrate, and concentrate under reduced pressure to a ratio of 1:2 (i.e., 1g of medicinal material corresponds to 2g of extract) to obtain Borage extract 4.

[0046] Example 5

[0047] Weigh a certain amount of Borage leaf powder, add 20 times the volume of ethanol, the ethanol concentration is 50%, ultrasonicate at room temperature for 30 minutes, repeat the above operation twice, combine the filtrate, and concentrate under reduced pressure to a ratio of 1:2 (i.e., 1g of medicinal material corresponds to 2g of extract) to obtain Borage extract 5.

[0048] Example 6

[0049] Weigh a certain amount of Borage leaf powder, add 20 times the volume of ethanol, the ethanol concentration is 60%, ultrasonicate at room temperature for 30 minutes, repeat the above operation twice, combine the filtrate, and concentrate under reduced pressure to a ratio of 1:2 (i.e. 1g of medicinal material corresponds to 2g of extract) to obtain Borage extract 6.

[0050] Example 7

[0051] Weigh a certain amount of Borage leaf powder, add 20 times the volume of ethanol, the ethanol concentration is 70%, ultrasonicate at room temperature for 30 minutes, repeat the above operation twice, combine the filtrate, and concentrate under reduced pressure to a ratio of 1:2 (i.e., 1g of medicinal material corresponds to 2g of extract) to obtain Borage extract 7.

[0052] Example 8

[0053] Weigh a certain amount of Borage leaf powder, add 20 times the volume of ethanol, the ethanol concentration is 80%, ultrasonicate at room temperature for 30 minutes, repeat the above operation twice, combine the filtrate, and concentrate under reduced pressure to a ratio of 1:2 (i.e., 1g of medicinal material corresponds to 2g of extract) to obtain Borage extract 8.

[0054] Example 9

[0055] Weigh a certain amount of Borage leaf powder, add 20 times the volume of ethanol, the ethanol concentration is 90%, ultrasonicate at room temperature for 30 minutes, repeat the above operation twice, combine the filtrate, and concentrate under reduced pressure to a ratio of 1:2 (i.e., 1g of medicinal material corresponds to 2g of extract) to obtain Borage extract 9.

[0056] Example 10

[0057] A certain amount of Borage leaf powder was weighed and added with 20 times the volume of ethanol to a concentration of 100%, and ultrasonic treatment was performed at room temperature for 30 minutes. The above operation was repeated twice, and the filtrates were combined and concentrated under reduced pressure to a ratio of 1:2 (i.e., 1 g of medicinal material corresponds to 2 g of extract) to obtain Borage extract 10.

[0058] Example 11

[0059] A certain amount of Borage leaf powder was weighed and added with 20 times the volume of methanol, with the methanol concentration being 100%. Ultrasonic treatment was performed at room temperature for 30 minutes. The above operation was repeated twice. The filtrates were combined and concentrated under reduced pressure to a ratio of 1:2 (i.e., 1 g of medicinal material corresponds to 2 g of extract) to obtain Borage extract 11.

[0060] Test Example 1

[0061] This test example tests the effects of borage extracts obtained by different extraction methods on the growth of pakchoy seedlings by pot culture. In this test example, the borage extract is used by foliar spraying.

[0062] 1. Test method

[0063] (1) Test samples

[0064] Ethanol borage extract (prepared in Examples 1 to 10) and methanol borage extract (prepared in Example 11) were prepared with pure water to a 1% stock solution (i.e., 10,000 ppm), filtered and sterilized, and then diluted and applied according to the experimental design.

[0065] (2) Experimental design

[0066] The experiment included 34 treatments in total, with 8 replicates per group. See Table 1 for details.

[0067] Table 1 Experimental design

[0068] (3) Test process

[0069] 1) Preparation of pakchoy test materials

[0070] Weigh equal weights (400g) of vermiculite and coconut coir sterilized at 121°C for 20 minutes, mix them in a 2:1 ratio, and pot them for later use. Soak pakchoy seeds in 5% sodium hypochlorite solution for 10 minutes, rinse them 5-6 times with water, and then sow them. When pakchoy seedlings have three leaves and a heart, select uniform, pest-free seedlings for planting and set aside.

[0071] 2) Preparation of pharmaceutical preparations

[0072] The 1% mother solution prepared by extracting and concentrating Borage extract with different solvents was diluted 200 times, 1000 times, and 5000 times respectively to obtain 50ppm, 10ppm, and 2ppm agents.

[0073] 3) Experimental treatment

[0074] Uniform, pest-free, plump pakchoy seedlings were selected and transplanted into pots, with one plant per pot and eight replicates per treatment. The seedlings were allowed to harden for three days, resulting in a total of 272 pots. After hardening, each germination box was watered with 50 mL of the pesticide solution according to the experimental design, while the control was watered with plain water. Indicators were assessed seven days after treatment.

[0075] (4) Detection indicators

[0076] The test indicators include: chlorophyll, leaf area, aboveground fresh weight, and aboveground dry weight.

[0077] 2. Results Analysis

[0078] (1) Effects of Borage Extracts Extracted by Different Extraction Methods on SPAD Values ​​of Chinese Cabbage

[0079] As shown in Table 2, root application of different borage extracts increased the SPAD value of pakchoi leaves compared to the water control. Comparison revealed that the SPAD value of pakchoi treated with 50%-70% ethanol-extracted borage extract had a relatively high growth rate, reaching a maximum of 31.25%.

[0080] Table 2 Effects of different borage extracts on SPAD values ​​of pakchoy

[0081] (2) Effects of different borage extracts on the fresh weight and dry weight of the aboveground part of pakchoy

[0082] As shown in Tables 3 and 4, the aboveground fresh weight and aboveground dry weight of pakchoy showed the same trend. The biomass of pakchoy treated with different borage extracts increased to varying degrees compared with the water control. Among them, the aboveground fresh weight of pakchoy treated with 60% ethanol borage extract was higher than that of other treatments at all three concentrations, with the highest growth rate reaching 30.33%. The growth rate of aboveground dry weight was the highest under the 10 ppm 60% ethanol borage extract treatment, reaching 27.52%.

[0083] Table 3 Effects of different borage extracts on the fresh weight of aboveground parts of Chinese cabbage

[0084] Table 4 Effects of different borage extracts on the dry weight of the aboveground part of pakchoy

[0085] (3) Effects of different borage extracts on the leaf area of ​​pakchoy

[0086] As shown in Table 5, after treatment with different borage extracts, the leaf area of ​​pakchoy leaves increased significantly. Under the treatment with 50%-60% ethanol extract, the leaf area growth rate increased significantly compared with other treatments. Among them, under the treatment with 50% ethanol extract 10ppm, the leaf area growth rate reached 38.00%. Secondly, under the treatment with 60% ethanol extract, the leaf area growth rates at different concentrations reached 33.33%, 36.67% and 29.98%, respectively.

[0087] Table 5 Effects of different borage extracts on the leaf area of ​​Chinese cabbage

[0088] In summary, Borage extract extracted with different alcohol solvents has a promoting effect on the growth of Chinese cabbage. The SPAD value, leaf area and biomass of Chinese cabbage leaves are significantly increased. Among them, after 50%-60% ethanol and ultrasonic solvent extraction, the growth-promoting effect of Borage extract is more prominent, and the performance is better under the 10ppm treatment.

[0089] Test Example 2

[0090] This test example tests the effects of three ethanol borage extracts that performed well in Test Example 1 on corn seed germination and seedling growth by pot culture. In this test example, the borage extracts were used for seed soaking.

[0091] 1. Test method

[0092] (1) Test samples

[0093] The ethanol borage extract (prepared in Examples 4, 6, and 8) was prepared with pure water to a 1% (i.e., 10,000 ppm) mother solution, which was then filtered and sterilized and then diluted and applied according to the experimental design.

[0094] (2) Experimental design

[0095] The experiment included 10 treatments in total, with 8 replicates per group. See Table 6 for details.

[0096] Table 6 Experimental design

[0097] (3) Test steps:

[0098] 1) Culture medium preparation

[0099] Weigh an equal weight (350 g) of vermiculite sterilized with high-pressure steam at 121° C. for 20 minutes and place it in pots for later use, for a total of 80 pots. Add 200 mL of clean water to each pot and stir thoroughly for later use.

[0100] 2) Seed soaking treatment

[0101] The prepared Borage extract stock solution was diluted 5000-fold, 1000-fold, and 200-fold, respectively, to produce seed soaking solutions with concentrations of 2 ppm, 10 ppm, and 50 ppm. For each treatment, 100 g of corn seeds were placed in an incubator. 200 mL of the seed soaking solution was added to the seed-containing incubator and mixed thoroughly with the seeds, completely immersing them in the solution. The seeds were then treated at room temperature for 24 hours. After 24 hours of soaking, the corn seeds were rinsed with clean water and sown into the culture medium, with 20 seeds per pot, for a total of 100 corn seeds per treatment. The results were evaluated after 5 days.

[0102] (4) Survey indicators

[0103] The test survey indicators include: germination rate, root length, root fresh weight, plant height, and plant fresh weight.

[0104] 2. Results Analysis

[0105] (1) Effects of soaking seeds with different borage extracts on corn germination rate

[0106] As shown in Table 7, the germination rate of corn increased after soaking the seeds with 40%-80% ethanol-extracted borage extract at 2, 10, and 50 ppm. The best performance was achieved with 10 ppm of the extract, and the highest germination rate growth rate reached 33.33%.

[0107] Table 7 Effect of Borage Extract Seed Soaking on Corn Germination Rate

[0108] (2) Effects of soaking seeds with different Borage extracts on the growth of corn seedlings

[0109] As shown in Tables 8 and 9, the effects of different Borage extracts on the growth of the aboveground part and root system of corn showed the same trend, and they all had obvious growth-promoting effects at different concentrations.

[0110] Overall, after treatment with various borage extracts, the performance of various indicators at 10ppm was better, with the highest growth rates of various indicators reaching 17.48%, 34.81%, 23.32%, and 17.57%, respectively; and the best performance was achieved under 60% ethanol extraction conditions.

[0111] Table 8 Effects of Borage Extract Treatment on Root Length and Plant Height Changes of Corn

[0112] Table 9 Effects of Borage Extract Treatment on Changes in Root Weight and Plant Weight of Corn

[0113] Test Example 3

[0114] This test example uses a pot culture method to test the effect of 60% ethanol borage extract, which performed well in Test Example 2, on rice seed germination and seedling growth when used as seed dressing under saline-alkali conditions. In this test example, the borage extract is used as seed dressing.

[0115] 1. Test method

[0116] (1) Test samples

[0117] 60% ethanol Borage extract (prepared in Example 6).

[0118] (2) Experimental design

[0119] The experiment included 4 treatments in total, with 5 replicates per group. See Table 10 for details.

[0120] Table 10 Experimental design

[0121] (3) Test steps:

[0122] 1) Culture medium preparation

[0123] Weigh an equal weight (100 g) of vermiculite sterilized with high-pressure steam at 121°C for 20 minutes and place it in pots for later use, a total of 20 pots. Add 80 mL of saline-alkali aqueous solution with a pH of 8.5 prepared with NaOH reagent to each pot, stir and mix thoroughly for later use.

[0124] 2) Seed dressing treatment: For each treatment, 100 g of rice seeds were placed in a ziplock bag. The required agent for 100 g of rice seeds was prepared according to the experimental design: 20 g / 2 L water / 100 kg of seeds, 100 g / 2 L water / 100 kg of seeds, and 500 g / 2 L water / 100 kg of seeds. The agent was thoroughly mixed with the seeds to completely cover the seed surface with the agent solution and treated at room temperature for 24 h.

[0125] 3) 24 hours after seed dressing, the rice seeds were sown into the spare culture medium according to the experimental design, 30 seeds were sown per pot, that is, 150 corn seeds were sown per treatment. The experimental results were investigated after 4-5 days.

[0126] (4) Survey indicators

[0127] The test survey indicators include: germination rate, plant height and plant weight.

[0128] 2. Results Analysis

[0129] (1) Effects of Borage Extract Seed Treatment on Rice Seed Germination

[0130] As shown in Table 11, under saline-alkali conditions, the germination rate of rice seeds treated with clear water was only 55.0%, while the germination rate of rice seeds treated with Borage extract increased significantly, with the highest germination rate reaching 91.67% and the highest growth rate reaching 66.67%.

[0131] Table 11 Effect of Borage Extract Seed Dressing on Rice Germination Rate under Saline-Alkali Conditions

[0132] Saline-alkali conditions inhibited the germination and growth of rice, and the rice seedlings grew slowly. Under the clear water treatment, the length of the rice seedlings was only 2.01 cm, and the average weight of a single plant was only 24.4 mg; after treatment with borage extract, the length of the rice seedlings reached 4.71 cm, the highest growth rate was 134.33%, and the highest growth rate of plant weight was 108.20%. This shows that borage extract seed dressing can promote the rapid growth of rice seedlings on the basis of promoting normal rice germination, thereby alleviating the impact of the saline-alkali environment on rice growth.

[0133] Table 12 Effects of Borage Extract Seed Dressing on Rice Seedling Growth under Saline-Alkali Conditions

[0134] Test Example 4

[0135] This test example uses a potted culture method to test the effect of 60% ethanol borage extract, which performed well in Test Example 2, on the growth of pepper seedlings under foliar spray treatment. In this test example, the borage extract is used by foliar spraying.

[0136] 1. Test method

[0137] (1) Test samples

[0138] 60% ethanol Borage extract (prepared in Example 6) was prepared into a 0.1% (ie, 1000 ppm) stock solution, filtered and sterilized, and then diluted and applied according to the experimental design.

[0139] (2) Experimental design

[0140] The experiment included 4 treatments in total, with 8 replicates per group. See Table 13 for details.

[0141] Table 13 Experimental design

[0142] (3) Test steps:

[0143] 1) Culture medium preparation

[0144] Sterilize vermiculite and coconut coir with high-pressure steam at 121℃ for 20 minutes, mix them evenly in a ratio of 2:1, and put them into pots for later use.

[0145] 2) After the pepper seeds are soaked and disinfected in 5% sodium hypochlorite solution for 10 minutes, they are washed with water 5-6 times and then sown; when the pepper seedlings grow to 4-5 leaves, the seedlings of uniform size and free of pests and diseases are selected for planting.

[0146] 3) Preparation of pharmaceutical preparations

[0147] The 60% ethanol Borage extract stock solution was diluted 5000 times, 1000 times, and 200 times to obtain 2ppm, 10ppm, and 50ppm of the drug.

[0148] 4) Experimental treatment

[0149] Uniformly sized, pest-free pepper seedlings were transplanted into pots, with one plant per pot and eight replicates per treatment, for a total of 32 pots. After three days of acclimatization, the prepared pesticides were sprayed according to the experimental design, covering the pepper leaves evenly but without dripping. A control was sprayed with plain water. Indicators were assessed seven days after treatment.

[0150] (4) Detection indicators

[0151] The test indicators include: chlorophyll, number of leaves, root length, plant height, root weight and plant weight.

[0152] 2. Results Analysis

[0153] (1) Effects of Borage Extract Treatment on Pepper Leaf Growth

[0154] As shown in Table 14, under the clean water treatment, the SPAD value of the pepper leaves was 41.23, while after the borage extract treatment, the SPAD value of the pepper leaves reached 48.55 at a concentration of 2 ppm, with a growth rate of 17.75%; after the borage extract treatment, the number of pepper leaves also increased to varying degrees, and under the 10 ppm treatment, the growth rate reached 9.62%.

[0155] Table 14 Effects of Borage Extract Treatment on Pepper Leaf Growth

[0156] (2) Effects of Borage Extract Treatment on the Growth of Pepper Seedlings

[0157] As shown in Table 15, after the leaves were treated with Borage extract, the growth of the aboveground part and root system of peppers were promoted to varying degrees, with the highest growth rates reaching 32.87%, 19.76%, 34.02%, and 23.54%, respectively. Among them, the performance was better under the 10ppm treatment.

[0158] Table 15 Effect of Borage Extract Treatment on Growth of Pepper Seedlings

[0159] Overall, foliar spraying of Borage extract can promote higher chlorophyll content in pepper seedlings and faster leaf growth, while also promoting plant height and root growth of pepper seedlings, and performs better under a concentration of 10 ppm.

[0160] Test Example 5

[0161] This test example tests the effect of 60% ethanol Borage extract, which performed well in Test Example 2, on the growth of cucumber seedlings under low temperature conditions by foliar spraying using a potted culture method. In this test example, the Borage extract was used by foliar spraying.

[0162] 1. Test method

[0163] (1) Test samples

[0164] 60% ethanol Borage extract (prepared in Example 6) was prepared into a 0.1% (ie, 1000 ppm) stock solution, filtered and sterilized, and then diluted and applied according to the experimental design.

[0165] (2) Experimental design

[0166] The experiment included 4 treatments in total, with 8 repetitions per group. See Table 16 for details.

[0167] Table 16 Experimental design

[0168] (3) Test steps:

[0169] 1) Preparation of culture medium: Vermiculite and coconut coir sterilized with high-pressure steam at 121°C for 20 minutes are mixed in a ratio of 2:1 and placed in a pot for later use.

[0170] 2) Material preparation: Cucumber seeds were disinfected by soaking in 5% sodium hypochlorite solution for 10 minutes, then washed 5-6 times with water before sowing. When the cucumber seedlings grew to the point where the first true leaf was fully expanded, seedlings of uniform size and free of pests and diseases were selected for later use.

[0171] 3) Preparation of pharmaceutical preparation: dilute the composite extract mother solution 5000 times, 1000 times, and 200 times to obtain pharmaceutical preparations of 2 ppm, 10 ppm, and 50 ppm.

[0172] 4) Experimental treatment

[0173] Cucumber seedlings of uniform size, free of pests and diseases, were selected and transplanted into pots, with one plant per pot and eight replicates per treatment, for a total of 32 pots. Each treatment was sprayed with the prepared pesticide according to the experimental design until the pakchoi leaves were evenly covered with the solution, but the solution did not drip off. The control was sprayed with water. Twenty-four hours after the spraying treatment, the seedlings were placed in a low-temperature environment at 4°C for 24 hours. The cucumbers were then observed for wilting and their water loss rate was measured.

[0174] (4) Detection indicators

[0175] The test indicator is the water loss rate of cucumber leaves.

[0176] The first true leaf of the cucumber after low temperature treatment was removed and weighed, and the leaf weight A was recorded. Then, the leaf was placed in clean water for 12 hours to allow the leaf to fully absorb the water to saturation, and the leaf weight after water absorption B was recorded.

[0177] Leaf water loss rate % = (BA) / B * 100%

[0178] 2. Results Analysis

[0179] After treatment at low temperatures (4°C), cucumber leaves in the water-treated group showed severe wilting, while those treated with Borage extract showed less wilting, with no noticeable curling or softening. The water loss rate of cucumber leaves under low temperatures was used to quantitatively measure the cold tolerance, with lower water loss rates indicating greater cold tolerance.

[0180] The water loss rate of cucumber leaves was measured and it was found that after treatment with Borage extract, the water loss rate of cucumber leaves was the lowest at 26.34%, which was 36.27% lower than that of the water treatment, and was mainly manifested under the 10 ppm concentration treatment.

[0181] Table 17 Effect of Borage Extract Treatment on Water Loss Rate of Cucumber Leaf Spray under Low Temperature Condition

[0182] Test Example 6

[0183] It can be seen from Test Examples 1-5 that borage extract, under 60% ethanol extraction conditions, has a significant effect of promoting growth and alleviating adverse damage on corn seed germination, rice seed germination under saline-alkali conditions, cucumber leaf spray growth under low temperature conditions, and pepper seedling growth. Therefore, 60% ethanol borage extract was used with surfactants such as thickeners and emulsifiers to trial-produce a borage extract preparation to verify the effect of a stable preparation on crop growth.

[0184] 1. Trial preparation of Borage extract preparation

[0185] The formulation includes: Borage extract, thickener - xanthan gum, emulsifier - alkyl glycoside, defoamer and water, which are compounded and trial-produced in different proportions. The specific formulation design table is as follows:

[0186] Table 18 Borage extract preparation trial formula

[0187] 2. Stability investigation of trial formula

[0188] After trial production of each formulation, the samples were subjected to high temperature (54°C) and low temperature (0°C) storage stability tests, and the formulation stability investigation was conducted after the samples were stored in hot and cold conditions for 2 weeks.

[0189] The results showed that a small amount of precipitation occurred in formulation 1 under hot storage conditions; the formulation 2 sample showed obvious stratification without the addition of emulsifier, and had poor dispersibility in water; the formulation 3 sample had a lot of foam during the dissolution process, and the formulation 4 preparation sample was a homogeneous stable liquid with good dispersibility.

[0190] Therefore, Formulation 4 was preferred as the trial formulation of Borage extract.

[0191] 3. Formulation trial production process

[0192] Borage extract, thickener, emulsifier, defoamer and water were prepared in a ratio of 20:10:2:0.5:67.5. The specific process operation method is as follows:

[0193] ① Weigh 2% xanthan gum and 85.5% water and shear until the xanthan gum is completely dissolved and dispersed to form a uniform liquid;

[0194] ② The liquid in ① was stirred at 55°C, and 20% of the borage extract was accurately weighed and added to a stirred tank. The mixture was stirred at 55°C for 30 minutes until the liquid was homogeneous.

[0195] ③ Weigh 2% of alkyl glycoside and add it to the above ② homogeneous liquid, stir for 3 minutes; then add 0.5% of defoaming agent and stir for 3 minutes to complete the trial preparation of the formula sample.

[0196] 4. Test results of Borage extract preparations

[0197] The basic physicochemical properties and technical indicators of Borago extract preparations are shown in Table 19:

[0198] Table 19 Test results of Borage extract preparation

[0199] After forming a stable sample of the borage extract preparation, the preparation is applied to biological test effect verification to clarify its application effect.

[0200] Test Example 7

[0201] This test example tests the effect of the borage extract preparation in Test Example 6 combined with 0.0075% brassinosteroid foliar spray on tomato growth and fruit quality by pot culture. In this test example, the borage extract preparation is applied by foliar spraying.

[0202] 1. Test method

[0203] (1) Test samples

[0204] In Experimental Example 6, the borage extract preparation was prepared in an amount of 20% (ie, the borage extract concentration was 200,000 ppm). The preparation was sterilized by filtration and then diluted and applied according to the experimental design.

[0205] (2) Experimental design

[0206] The experiment included 4 treatments in total, with 8 repetitions per group. See Table 20 for details.

[0207] Table 20 Experimental design

[0208] (3) Experimental treatment:

[0209] 1) Culture medium preparation

[0210] Vermiculite and coconut coir sterilized with high-pressure steam at 121℃ for 20 minutes are mixed evenly in a ratio of 2:1 and placed in a pot for later use.

[0211] 2) Material preparation: Tomato seeds were disinfected by soaking in 5% sodium hypochlorite solution for 10 minutes, then rinsed with water and sown. When the tomato seedlings grew to 4-5 leaves, seedlings of uniform size and free of pests and diseases were selected for planting.

[0212] 3) Preparation of pharmaceutical preparations

[0213] The actual spraying concentrations were as follows: diluting the borage extract preparation 4000 times and diluting 0.0075% brassinolide 3000 times.

[0214] 4) Experimental treatment

[0215] Uniformly sized, pest-free tomato seedlings were transplanted into pots, with one plant per pot and eight replicates per treatment, for a total of 32 pots. The pesticides were sprayed according to the experimental design during the seedling stage (7-8 leaves), pre-flowering, and fruiting stages. At harvest, leaf growth and plant height were assessed, and tomato fruit yield was measured.

[0216] (5) Detection indicators

[0217] The test indicators include: chlorophyll, plant height, number of tomato flowers, number of fruits, fruit reddening rate and average single plant yield.

[0218] 2. Results Analysis

[0219] (1) Effect of spraying Borage extract preparation on tomato growth

[0220] As can be seen from Table 21, after treatment with the plant growth regulator brassinolide and borage extract preparations, the SPAD value of tomato leaves and the tomato plant height increased significantly compared with the water control, with the growth rate ranging from 7.37% to 11.22% and 9.20% to 18.99%; compared with the single-agent treatment, the SPAD value of tomato leaves and the plant height increased to varying degrees after treatment with the combination of brassinolide and borage extract preparations, with the highest increase of 5.07% in SPAD value and 11.14% in plant height.

[0221] Table 21 Effects of Borage Extract Preparation Treatment on Tomato Growth

[0222] (2) Effects of Borage Extract Preparation on Tomato Fruit Growth

[0223] As shown in Table 22, after treatment with the plant growth regulator brassinolide and borage extract, the average number of flowers and fruits per tomato plant increased to varying degrees compared with the water treatment, and the increase was greater under the borage extract treatment; similarly, the red-turning rate of tomato fruits and the average yield per plant also increased to varying degrees, with the highest growth rates reaching 24.52% and 24.66% respectively.

[0224] Compared with single-agent treatment, the number of tomato flowers, number of fruits, red-turning rate and single-plant yield increased to varying degrees after the combination treatment of brassinolide and borage extract preparation, indicating that the borage extract preparation combined with the regulator treatment can promote tomato fruit growth and fruit setting, accelerate the color change of tomato fruit, and increase tomato fruit yield.

[0225] Table 22 Effects of Borage Extract Preparation Treatment on Tomato Fruit Growth

[0226] Test Example 8

[0227] The effect of borage extract preparation combined with potassium dihydrogen phosphate foliar spray on wheat yield and quality was tested.

[0228] 1. Test method

[0229] (1) Test sample preparation

[0230] The borage extract preparation prepared as described in Experimental Example 6, wherein the borage extract was added in an amount of 20% (ie, the concentration of the active substance was 200,000 ppm), was filter-sterilized, and then diluted and applied according to the experimental design.

[0231] (2) Experimental design

[0232] Table 23 Experimental design

[0233] Each treatment was set up with 3 replicates, and the size of each replicate plot was 2m*3m, for a total of 12 treatment plots.

[0234] (3) Experimental treatment

[0235] During the heading and filling stages of wheat, pesticides were sprayed according to the experimental design. The thousand-grain weight and plot yield of wheat were investigated at harvest, and samples were taken for quality testing.

[0236] 2. Results Analysis

[0237] Borage extract, applied at different wheat growth stages, boosted thousand-kernel weight and yield. Compared to the water control, thousand-kernel weight increased by 13.20% and yield per square plot increased by 11.13%. When Borage extract was combined with potassium dihydrogen phosphate foliar fertilizer, wheat yield was significantly improved, with thousand-kernel weight increasing from 44.6g to 45.7g and yield per square plot increasing by 3.2g.

[0238] Table 24 Effect of Borage Extract Preparation on Wheat Yield

[0239] Testing of wheat grain quality across different treatments revealed significant increases in protein and starch content after borage extract treatment compared to water treatment, reaching 16.55% and 15.41%, respectively. Foliar spraying of borage extract combined with potassium dihydrogen phosphate also significantly improved wheat grain quality, with protein content increasing by 3.37% and starch content by 3.88%, compared to treatment with borage extract alone.

[0240] Overall, foliar spraying of Borage extract preparation can significantly promote wheat yield and quality improvement. When combined with potassium dihydrogen phosphate treatment, it has a synergistic effect on improving wheat quality.

[0241] Table 25 Effect of Borage Extract Preparation on Wheat Quality

[0242] Test Example 9

[0243] The effects of foliar spraying of borage extract preparation on increasing yield and improving quality of pakchoy were tested.

[0244] 1. Test method

[0245] (1) Test sample preparation

[0246] In Experimental Example 6, the Borage extract preparation was prepared in an amount of 20% (ie, the concentration of the active substance was 200,000 ppm). The preparation was sterilized by filtration and then diluted and applied according to the experimental design.

[0247] (2) Experimental design

[0248] Table 26 Experimental design

[0249] (3) Experimental treatment

[0250] Pakchoy grown in the open field was selected as the experimental crop. Borage extract preparations were sprayed according to the experimental design at the seedling stage (three leaves and one heart) and the rapid growth stage (6-7 leaves). The changes in leaf color and yield of the pakchoy were investigated at harvest, and samples were taken for determination of protein and vitamin C nutritional quality.

[0251] 2. Results Analysis

[0252] After treatment with different concentrations of borage extract, the leaves of Chinese cabbage became darker green, the SPAD value increased by up to 21.42%, and the yield per square plot increased by up to 35.16% compared with the water control treatment.

[0253] Table 27 Effect of Borage Extract Preparation Leaf Spray on the Growth of Chinese Cabbage

[0254] After foliar spraying with Borage extract, the vitamin C and protein content of Chinese cabbage increased to varying degrees. Among them, under the treatment with a concentration of 250ppm, the quality improvement effect was better, with the vitamin C content increasing by 23.72% and the protein content increasing by 9.74%.

[0255] Table 28 Effect of leaf spraying of borage extract preparation on the quality of pakchoy

[0256] Overall, foliar spraying of Borage extract preparation can promote the growth of Chinese cabbage leaves and increase the yield of Chinese cabbage. At the same time, it can improve the quality of the edible parts of Chinese cabbage and increase the content of vitamin C and protein.

[0257] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. Application of Borage extract in promoting plant growth and / or regulating plant stress resistance.

2. The use according to claim 1, characterized in that: The borage extract can promote seed germination and / or the growth of one or more of roots, stems, leaves, or flowers and fruits.

3. The use according to claim 1, characterized in that: The stress resistance is at least one of salt resistance, cold resistance, drought resistance and high temperature resistance.

4. The use according to claim 1, characterized in that: The Borage extract is an alcohol extract; further, the alcohol solution is selected from methanol or ethanol.

5. The use according to claim 4, characterized in that: The alcohol solution is ethanol; further, the concentration of ethanol is 10% to 100%; further, the concentration of ethanol is 40% to 80%.

6. The use according to claim 4, characterized in that: The mass ratio of the borage to the alcohol solution is 1-5:15-25, preferably 1:

20.

7. The use according to claim 1, characterized in that: The concentration of the Borage extract is 0.001 ppm to 100 ppm, preferably 2 to 50 ppm.

8. The use according to claim 1, characterized in that: When in use, the borage extract is made into an agricultural product, which also includes auxiliary materials, such as one or more of a thickener, a defoamer, a dispersant, a wetting agent, a binder, an emulsifier, a stabilizer and a solvent.

9. The use according to claim 8, characterized in that: The dosage form of the product is powder, granule, aqueous solution, mother liquid or mother powder.

10. The application according to claim 9, characterized in that: The agricultural products made from Borago extract are used in combination with plant regulators, foliar fertilizers, water-soluble fertilizers, compound fertilizers and pesticides.

11. The use according to claim 10, characterized in that: When used, a single dose of Borage extract or the prepared agricultural product is used to treat seeds, spray leaves or irrigate roots.

12. A biostimulant, characterized in that: It contains Borage extract as its active ingredient.

13. The use or biostimulant according to any one of claims 1 to 12, characterized in that: The plants include tobacco, corn, rice, cucumber, lettuce, wheat, pepper, Chinese cabbage, lettuce, bok choy, tomato, citrus, kiwi, cherry, pear and apple.

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