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

Abstract

The present invention belongs to the field of agricultural technology, and specifically relates to the application of borage extract in promoting plant growth and / or regulating plant stress resistance. At present, borage extract has only been reported to be used in the medical field, and has not been involved in agriculture. The present invention provides a new application of borage extract, specifically using it as a biostimulant to promote plant growth and enhance plant stress resistance.

Description

Technical Field

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

[0002] Borage is a plant of the Boraginaceae family. It is a strong, annual herb. It is an annual herb that can reach 120 cm in height. The whole plant is covered with rough, gray-white prickles. The stem is cylindrical, about 11 / 2 feet high, and has branches. Its branches are hollow and rich in water. The leaves are dark green, wrinkled, and grow alternately on the stem; they are about 3 inches long and 11 / 2 inches wide; the leaves are rigidly and gradually symmetrically contracted from bottom to top into an ellipsoidal shape; the burrs covering the upper and lower leaves and the stem have very small rounded heads at the top, making the outer burr edges of the ellipsoidal body formed by the leaves continuously wavy. Its fruit contains four dark brown nuts. The leaves are alternate and rough like cucumber leaves, with petioles. The flowering period is July, the flowers are blue, and they are in sparse, leafy cymes with long petioles. Small nuts, smooth or with nipple-like protrusions.

[0003] Borago and / or Borago extracts have certain nutritional value and physiological functions, and are often used in the medical field, mainly including: (1) prevention and treatment of cardiovascular diseases; Borago and / or Borago extracts can inhibit angiotensin synthesis to lower blood pressure, and reduce serum cholesterol and triglyceride levels; (2) treatment of diabetes; Borago and / or Borago 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 Borago extracts are used to promote plant growth. Summary of the invention

[0004] The present invention discovers for the first time that Borago extract not only promotes plant growth, but also enhances plant stress resistance, thereby expanding the application scope of Borago extract and providing a new option for regulating plant growth in agriculture.

[0005] The invention provides application of Borage extract in promoting plant growth and / or regulating plant stress resistance.

[0006] 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 thickness, plant height, leaf width, leaf length, leaf number, leaf area, biomass, chlorophyll content, yield, etc.

[0007] 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.

[0008] In the present invention, the stress resistance includes but is not limited to salt-alkali resistance, cold resistance, drought resistance, and high temperature resistance.

[0009] In the present invention, the Borage extract is an alcohol extract.

[0010] In the present invention, the alcohol solution is selected from methanol, ethanol and propanol.

[0011] In a specific embodiment of the present invention, the alcohol solution is ethanol or methanol.

[0012] In the present invention, the ethanol concentration or the 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%, can also be selected from 30%, 40%, 50%, 60%, 70%, 80% and the like.

[0013] Furthermore, the concentration of the ethanol is 40% to 80%.

[0014] In the present invention, the mass ratio of the borage to the ethanol is 1-5:15-25.

[0015] In a specific embodiment of the present invention, the mass ratio of Borage to ethanol is 1:20.

[0016] When mass ratio or other value or parameter is represented by range, preferred range, or a series of upper preferred values ​​and lower preferred values, 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, regardless of whether the range is disclosed separately. For example, when disclosing range "1 to 5", the described range should be interpreted as including range "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, the range is intended to include its end value and all integers and fractions within the range.

[0017] 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.

[0018] The invention provides a biostimulant which takes borage extract as an active ingredient.

[0019] The "biostimulants" mentioned in the present invention include substances and / or microorganisms, which, when applied to plants or the rhizosphere, function to stimulate natural processes to promote and / or inhibit nutrient absorption, increase and / or reduce nutrient conversion efficiency, abiotic tolerance and crop yield.

[0020] In the present invention, the borage extract is a natural biostimulant.

[0021] 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.

[0022] 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.

[0023] 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.).

[0024] In the product of the present invention, the borage extract can be directly used as a single dose. In order to make it stable and convenient for transportation and storage, it can be made into an agricultural product, for example, by adding auxiliary materials to make a corresponding dosage form. The auxiliary materials can be conventional auxiliary materials in the field, such as: thickeners, defoamers, dispersants, wetting agents, binders, emulsifiers, stabilizers, solvents, etc.

[0025] On the other hand, the Borago 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, pesticides, etc.

[0026] In the present invention, the dosage form of the agricultural product includes but is not limited to powder, granule, aqueous solution, mother solution or mother powder.

[0027] In the present invention, when used, the single agent or agricultural product prepared from the borage extract is used to treat seeds, spray leaves or irrigate roots.

[0028] In the present invention, when the Borago extract or its product is prepared into a solution for promoting seed germination or plant growth, the concentration of the Borago extract in the solution can be selected according to actual needs.

[0029] 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,

[0030] 1.0ppm, 2.0ppm, 3.0ppm, 10ppm, 20ppm, 30ppm and so on.

[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 the seeds to be germinated with borage extract solution.

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

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

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

[0037] The technical solution of the present invention is described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0038] Example 1

[0039] Weigh a certain amount of Borago leaves and grind them into powder. Add 20 times the volume of ethanol with an ethanol concentration of 10%, perform ultrasonic treatment 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 Borago extract 1.

[0040] Example 2

[0041] Weigh a certain amount of Borago leaves and grind them into powder. Add 20 times the volume of ethanol with an ethanol concentration of 20%, perform ultrasonic treatment 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 Borago extract 2.

[0042] Example 3

[0043] Weigh a certain amount of Borago leaves and grind them into powder. Add 20 times the volume of ethanol with an ethanol concentration of 30%, perform ultrasonic treatment 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 Borago extract 3.

[0044] Example 4

[0045] Weigh a certain amount of Borago leaves and grind them into powder. Add 20 times the volume of ethanol with an ethanol concentration of 40%, perform ultrasonic treatment 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 Borago extract 4.

[0046] Example 5

[0047] Weigh a certain amount of Borago leaves and grind them into powder. Add 20 times the volume of ethanol with an ethanol concentration of 50%, perform ultrasonic treatment 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 Borago 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 for 30 minutes at room temperature, repeat the above operation twice, combine the filtrate, and concentrate under reduced pressure to 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 Borago leaves and grind them into powder. Add 20 times the volume of ethanol with an ethanol concentration of 70%, and perform ultrasonic treatment 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 Borago extract 7.

[0052] Example 8

[0053] Weigh a certain amount of Borago officinalis leaves and grind them into powder. Add 20 times the volume of ethanol with an ethanol concentration of 80%, perform ultrasonic treatment 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 Borago officinalis extract 8.

[0054] Example 9

[0055] Weigh a certain amount of Borago leaves and grind them into powder. Add 20 times the volume of ethanol with an ethanol concentration of 90%, perform ultrasonic treatment 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 Borago extract 9.

[0056] Example 10

[0057] Weigh a certain amount of Borago officinalis leaves and grind them into powder. Add 20 times the volume of ethanol, whose ethanol concentration is 100%, and ultrasonicate for 30 minutes at room temperature. Repeat the above operation twice, combine the filtrate, and concentrate under reduced pressure to 1:2 (i.e., 1 g of medicinal material corresponds to 2 g of extract) to obtain Borago officinalis extract 10.

[0058] Embodiment 11

[0059] Weigh a certain amount of Borago officinalis leaves and grind them into powder. Add 20 times the volume of methanol with a methanol concentration of 100%, perform ultrasonic treatment 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 Borago officinalis extract 11.

[0060] Test Example 1

[0061] In this test example, the effect of Borago extracts obtained by different extraction methods on the growth of Chinese cabbage seedlings was tested by pot culture. In this test example, the Borago extract was used by leaf spraying.

[0062] 1. Test methods

[0063] (1) Test samples

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

[0065] (2) Experimental design

[0066] There were a total of 34 treatments in the experiment, each repeated 8 times, see Table 1 for details.

[0067] Table 1 Experimental design

[0068]

[0069] (3) Test process

[0070] 1) Preparation of Chinese cabbage test materials

[0071] Weigh equal weight (400 g) of vermiculite and coconut bran sterilized with 121° C. high pressure steam for 20 minutes, mix them evenly in a ratio of 2:1, and pot them for later use. Soak the Chinese cabbage seeds in 5% sodium hypochlorite solution for 10 minutes, wash them with water 5-6 times, and then sow them; when the Chinese cabbage seedlings grow to three leaves and one heart, select seedlings of uniform size and free of pests and diseases for planting and set aside.

[0072] 2) Preparation of medicine

[0073] The 1% mother solution prepared by extracting and concentrating Borago officinalis with different solvents was diluted 200 times, 1000 times, and 5000 times respectively to obtain 50 ppm, 10 ppm, and 2 ppm of the drug.

[0074] 3) Experimental treatment

[0075] Select the same size, pest-free and full pakchoi seedlings and transplant them into pots, one plant per pot, 8 replicates per treatment, and 3 days for seedling acclimatization, for a total of 272 pots. After seedling acclimatization, 50 mL of the drug solution was poured into each germination box according to the experimental design, and the control was poured with water. Indicators were investigated 7 days after treatment.

[0076] (4) Detection indicators

[0077] The test indicators include: chlorophyll, leaf area, aboveground fresh weight, and aboveground dry weight.

[0078] 2. Results Analysis

[0079] (1) Effects of Borage Extracts Extracted by Different Extraction Methods on SPAD Values ​​of Chinese Cabbage

[0080] As shown in Table 2, compared with the water control, the SPAD value of pakchoi leaves increased after root application of different borage extracts. Comparison found that under different concentrations, the SPAD value growth rate of pakchoi after treatment with 50%-70% ethanol-extracted borage extract was relatively high, reaching a maximum of 31.25%.

[0081] Table 2 Effects of different Borage extracts on SPAD values ​​of Chinese cabbage

[0082]

[0083] (2) Effects of different Borage extracts on the fresh weight and dry weight of the aboveground part of Chinese cabbage

[0084] As shown in Tables 3 and 4, the aboveground fresh weight and aboveground dry weight of Chinese cabbage showed the same trend. The biomass of Chinese cabbage treated with different borage extracts increased to varying degrees compared with the water control. Among them, the aboveground fresh weight of Chinese cabbage treated with 60% ethanol borage extract at three concentrations was higher than that of other treatments, with the highest growth rate reaching 30.33%. Under the treatment with 10ppm 60% ethanol borage extract, the growth rate of aboveground dry weight was the highest, reaching 27.52%.

[0085] Table 3 Effects of different Borage extracts on the fresh weight of aboveground parts of Chinese cabbage

[0086]

[0087] Table 4 Effects of different Borage extracts on the aboveground dry weight of Brassica chinensis

[0088]

[0089]

[0090] (3) Effects of different Borage extracts on the leaf area of ​​Chinese cabbage

[0091] As shown in Table 5, after treatment with different borage extracts, the leaf area of ​​Chinese cabbage 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 extraction 10ppm, the leaf area growth rate reached 38.00%, followed by 60% ethanol extraction treatment, the leaf area growth rates at different concentrations reached 33.33%, 36.67% and 29.98% respectively.

[0092] Table 5 Effects of different Borage extracts on the leaf area of ​​Chinese cabbage

[0093]

[0094] 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. 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.

[0095] Test Example 2

[0096] In this test example, the effects of the three ethanol Borago extracts that performed well in Test Example 1 on corn seed germination and seedling growth were tested by pot culture. In this test example, the Borago extracts were used for seed soaking.

[0097] 1. Test methods

[0098] (1) Test samples

[0099] Ethanol Borage Extract (prepared in Examples 4, 6, and 8) The above samples were all prepared with pure water to a 1% (ie, 10,000 ppm) mother solution, filtered and sterilized, and then diluted and applied according to the experimental design.

[0100] (2) Experimental design

[0101] There were 10 treatments in total, each repeated 8 times, see Table 6 for details.

[0102] Table 6 Experimental design

[0103]

[0104] (3) Test steps:

[0105] 1) Culture matrix preparation

[0106] 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.

[0107] 2) Seed soaking

[0108] Dilute the prepared Borage extract mother solution 5000 times, 1000 times, and 200 times to obtain 2ppm, 10ppm, and 50ppm soaking solutions. For each treatment, take 100g corn seeds and put them into the culture box. Take 200mL of soaking solution and add it to the culture box with seeds. Mix it with the seeds thoroughly so that the seeds are completely immersed in the solution. Treat at room temperature for 24 hours. After soaking for 24 hours, rinse the corn seeds with clean water and sow them into the culture matrix. Sow 20 seeds per pot, that is, sow 100 corn seeds for each treatment. Investigate the test results after 5 days.

[0109] (4) Survey indicators

[0110] The test survey indicators include: germination rate, root length, root fresh weight, plant height, and plant fresh weight.

[0111] 2. Results Analysis

[0112] (1) Effects of soaking seeds with different Borago extracts on corn germination rate

[0113] 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 extracts at different concentrations, with the highest germination rate growth rate reaching 33.33%.

[0114] Table 7 Effect of Borago extract soaking on corn germination rate

[0115]

[0116] (2) Effects of soaking seeds with different Borago extracts on the growth of corn seedlings

[0117] As shown in Tables 8 and 9, the effects of different Borago 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.

[0118] Overall, after treatment with various borage extracts, the performance of each index was better at 10ppm, with the highest growth rates of each index reaching 17.48%, 34.81%, 23.32%, and 17.57%, respectively; and the best performance was achieved under 60% ethanol extraction conditions.

[0119] Table 8 Effects of Borago extract treatment on root length and plant height of corn

[0120]

[0121] Table 9 Effects of Borago extract treatment on changes in root weight and plant weight of corn

[0122]

[0123] Test Example 3

[0124] This test example uses a pot culture method to test the effect of 60% ethanol Borago extract, which performed well in Test Example 2, on rice seed germination and seedling growth under saline-alkali conditions. In this test example, the Borago extract is used as seed dressing.

[0125] 1. Test methods

[0126] (1) Test samples

[0127] 60% ethanol Borage extract (prepared in Example 6).

[0128] (2) Experimental design

[0129] There were 4 treatments in total, each repeated 5 times, see Table 10 for details.

[0130] Table 10 Experimental design

[0131]

[0132] (3) Test steps:

[0133] 1) Culture matrix preparation

[0134] Weigh an equal weight (100 g) of vermiculite sterilized with high-pressure steam at 121°C for 20 minutes and put it into 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.

[0135] 2) Seed dressing treatment: For each treatment, 100 g of rice seeds were taken and put into a ziplock bag. According to the experimental design, 20 g / 2L water / 100 kg of seeds, 100 g / 2L water / 100 kg of seeds, and 500 g / 2L water / 100 kg of seeds were prepared. The required agent for 100 g of rice seeds was fully mixed with the seeds so that the surface of the seeds was completely covered with the agent, and the agent was treated at room temperature for 24 h.

[0136] 3) 24 hours after the rice seeds were mixed, they 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, and the experimental results were investigated after 4-5 days.

[0137] (4) Survey indicators

[0138] The test survey indicators include: germination rate, plant height and plant weight.

[0139] 2. Results Analysis

[0140] (1) Effects of Borage Extract Seed Treatment on Rice Seed Germination

[0141] 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 Borago extract increased significantly, with the highest germination rate reaching 91.67% and the highest growth rate reaching 66.67%.

[0142] Table 11 Effect of Borago extract seed dressing on rice germination rate under saline-alkali conditions

[0143]

[0144] Saline-alkali conditions inhibited the germination and growth of rice, and the rice seedlings grew slowly. Under the clean 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, and alleviate the impact of saline-alkali environment on rice growth.

[0145] Table 12 Effect of Borago extract seed dressing on rice seedling growth under saline-alkali conditions

[0146]

[0147]

[0148] Test Example 4

[0149] In this test example, potted culture was used to test the effect of 60% ethanol Borago extract, which performed well in Test Example 2, on the growth of pepper seedlings under foliar spray treatment. In this test example, the Borago extract was used by foliar spraying.

[0150] 1. Test methods

[0151] (1) Test samples

[0152] 60% ethanol Borago extract (prepared in Example 6) was prepared into a 0.1% (ie, 1000 ppm) stock solution, which was filtered and sterilized and then diluted and applied according to the experimental design.

[0153] (2) Experimental design

[0154] There were 4 treatment groups in total, each group was repeated 8 times, see Table 13 for details.

[0155] Table 13 Experimental design

[0156]

[0157] (3) Test steps:

[0158] 1) Culture matrix preparation

[0159] Sterilize vermiculite and coconut bran with high-pressure steam at 121℃ for 20 minutes, mix them evenly in a ratio of 2:1, and put them in a pot for later use.

[0160] 2) After the pepper seeds are soaked and disinfected in 5% sodium hypochlorite solution for 10 minutes, they are washed with water for 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.

[0161] 3) Preparation of medicine

[0162] The 60% ethanol Borago extract stock solution was diluted 5000 times, 1000 times, and 200 times to obtain 2ppm, 10ppm, and 50ppm agents.

[0163] 4) Experimental treatment

[0164] Pepper seedlings of uniform size and 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. After 3 days of seedling acclimatization, the prepared agent was sprayed on each treatment according to the experimental design until the pepper leaves were evenly covered with the liquid, but the liquid did not drip down. The control was sprayed with water. The indicators were investigated 7 days after the treatment.

[0165] (4) Detection indicators

[0166] The test indicators include: chlorophyll, number of leaves, root length, plant height, root weight and plant weight.

[0167] 2. Results Analysis

[0168] (1) Effects of Borago extract treatment on pepper leaf growth

[0169] It can be seen from Table 14 that 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 the 2ppm concentration treatment, 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 10ppm treatment, the growth rate reached 9.62%.

[0170] Table 14 Effect of Borago extract treatment on pepper leaf growth

[0171]

[0172] (2) Effects of Borago extract treatment on the growth of pepper seedlings

[0173] As shown in Table 15, after the leaves were treated with Borago 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. The performance was better under the 10ppm treatment.

[0174] Table 15 Effect of Borago extract treatment on the growth of pepper seedlings

[0175]

[0176] Overall, foliar spraying of Borage extract can promote higher chlorophyll content in pepper seedlings and faster leaf growth, while promoting plant height and root growth of pepper seedlings, and performs better under 10ppm concentration treatment.

[0177] Test Example 5

[0178] This test example tests the effect of 60% ethanol Borago extract, which performed well in Test Example 2, on the growth of cucumber seedlings by foliar spraying under low temperature conditions by potted cultivation. In this test example, the Borago extract was used by foliar spraying.

[0179] 1. Test methods

[0180] (1) Test samples

[0181] 60% ethanol Borago extract (prepared in Example 6) was prepared into a 0.1% (ie, 1000 ppm) stock solution, which was filtered and sterilized and then diluted and applied according to the experimental design.

[0182] (2) Experimental design

[0183] There were 4 treatment groups in total, each group was repeated 8 times, see Table 16 for details.

[0184] Table 16 Experimental design

[0185]

[0186] (3) Test steps:

[0187] 1) Preparation of culture medium: Vermiculite and coconut bran sterilized with high pressure steam at 121℃ for 20 minutes are mixed in a ratio of 2:1 and placed in pots for later use.

[0188] 2) Preparation of test materials: Cucumber seeds were soaked and disinfected in 5% sodium hypochlorite solution for 10 min, then washed with water for 5-6 times and sown; when the cucumber seedlings grew to the point where the first true leaf was fully unfolded, seedlings of uniform size and free of pests and diseases were selected for later use.

[0189] 3) Preparation of the drug: dilute the mother solution of the composite extract by 5000 times, 1000 times, and 200 times to obtain 2ppm, 10ppm, and 50ppm drugs.

[0190] 4) Experimental treatment

[0191] Select the cucumber seedlings of the same size, free of pests and diseases and transplant them into pots, with one plant per pot and eight replicates per treatment, for a total of 32 pots. According to the experimental design, each treatment was sprayed with the prepared agent until the leaves of the pakchoi were evenly covered with the liquid, but the liquid did not drip down. The control was sprayed with water. After 24 hours of spraying, the seedlings were placed in a low temperature environment of 4℃ for 24 hours, and then the wilting of the cucumber was observed and the water loss rate was measured.

[0192] (4) Detection indicators

[0193] The test indicator is the water loss rate of cucumber leaves.

[0194] 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 water to saturation, and the leaf weight after water absorption B was recorded.

[0195] Leaf water loss rate % = (BA) / B * 100%

[0196] 2. Results Analysis

[0197] After low temperature (4°C) treatment, it can be observed that the cucumber leaves in the water treatment group are severely wilted, while the cucumber leaves treated with Borago extract have a lower degree of wilting and no obvious curling and softening. The water loss rate of cucumber leaves under low temperature conditions is used to quantitatively calculate the anti-low temperature effect. The smaller the water loss rate of the leaves, the better the anti-low temperature effect.

[0198] 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 as low as 26.34%, which was 36.27% lower than that of the clean water treatment, and was mainly manifested under the 10ppm concentration treatment.

[0199] Table 17 Effect of Borago extract treatment on water loss rate of cucumber leaf spray under low temperature conditions

[0200]

[0201] Test Example 6

[0202] It can be seen from Test Examples 1-5 that Borago 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 Borago extract was used with surfactants such as thickeners and emulsifiers to trial-produce Borago extract preparations to verify the effect of stable preparations on crop growth.

[0203] 1. Trial preparation of Borago extract preparation

[0204] The formulation includes: Borago extract, thickener-xanthan gum, emulsifier-alkyl glycoside, defoamer and water, which are compounded and trial-produced according to different proportions. The specific formula design table is as follows:

[0205] Table 18 Borago extract preparation trial formula

[0206]

[0207] 2. Investigation on stability of trial formulation

[0208] 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.

[0209] The results showed that a small amount of precipitation occurred in Formulation 1 under hot storage conditions; in the absence of an emulsifier, the sample of Formulation 2 showed obvious stratification and had poor dispersibility in water; the sample of Formulation 3 had a lot of foam during the dissolution process, and the sample of Formulation 4 was a homogeneous stable liquid with good dispersibility.

[0210] Therefore, Formulation 4 is preferred as the trial formulation of Borage extract.

[0211] 3. Formulation trial production process

[0212] 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:

[0213] 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 for later use;

[0214] ② The liquid in ① is stirred at 55° C., and 20% of the Borago extract is accurately weighed and added into a stirring kettle, and stirred at 55° C. for 30 minutes until the liquid is homogeneous;

[0215] ③ Weigh 2% of alkyl polyglycoside 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 production of the formula sample.

[0216] 4. Test results of Borago extract preparations

[0217] The basic physicochemical properties and technical indicators of Borago extract preparation are shown in Table 19:

[0218] Table 19 Test results of Borago extract preparation

[0219]

[0220] After forming a stable sample of the borage extract preparation, the preparation is applied to biological test effect verification to clarify its application effect.

[0221] Test Example 7

[0222] This test example tests the effect of the borage extract preparation in Test Example 6 combined with the plant regulator 0.0075% brassinolide sterol foliar spray on tomato growth and fruit quality by pot culture. In this test example, the borage extract preparation is used by foliar spraying.

[0223] 1. Test methods

[0224] (1) Test samples

[0225] In Experimental Example 6, the Borago extract preparation was prepared in an amount of 20% (ie, the Borago extract concentration was 200,000 ppm). The preparation was sterilized by filtration and then diluted and applied according to the experimental design.

[0226] (2) Experimental design

[0227] There were 4 treatment groups in total, each group was repeated 8 times, see Table 20 for details.

[0228] Table 20 Experimental design

[0229]

[0230] (3) Experimental treatment:

[0231] 1) Culture matrix preparation

[0232] Vermiculite and coconut bran sterilized with high-pressure steam at 121℃ for 20 minutes are mixed in a ratio of 2:1 and placed in a pot for later use.

[0233] 2) Preparation of test materials: Tomato seeds were disinfected by soaking in 5% sodium hypochlorite solution for 10 min, then washed 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.

[0234] 3) Preparation of medicine

[0235] The actual spraying concentration is obtained by diluting the Borage extract preparation 4000 times and 0.0075% Brassinolide preparation 3000 times.

[0236] 4) Experimental treatment

[0237] Tomato seedlings of uniform size and 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. The pesticides were sprayed according to the experimental design during the tomato seedling stage (7-8 leaves), pre-flowering and fruiting stages. At harvest, the growth of tomato leaves and plant height were investigated, and the tomato fruit yield was measured.

[0238] (5) Testing indicators

[0239] The test indicators include: chlorophyll, plant height, number of tomato flowers, number of fruits, fruit turning red rate and average single plant yield.

[0240] 2. Results Analysis

[0241] (1) Effect of spraying Borago extract preparation on tomato growth

[0242] 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 clear water control, with growth rates ranging from 7.37%-11.22% and 9.20%-18.99%; compared with single-agent treatment, after treatment with the combination of brassinolide and borage extract preparations, the SPAD value of tomato leaves and the plant height increased to varying degrees, with the highest increase in SPAD value being 5.07% and the highest increase in plant height being 11.14%.

[0243] Table 21 Effect of Borage Extract Preparation Treatment on Tomato Growth

[0244]

[0245] (2) Effects of Borago extract preparation on tomato fruit growth

[0246] As can be seen from Table 22, after the treatment with the plant growth regulator brassinolide and borage extract preparations, the average number of flowers and fruits per tomato plant increased to varying degrees compared with the water treatment, and both showed a greater increase under the borage extract preparation treatment; similarly, the red fruit rate and average single plant yield of tomatoes also increased to varying degrees, with the highest growth rates reaching 24.52% and 24.66% respectively.

[0247] Compared with single-agent treatment, the number of tomato flowers, fruit set, red-turning rate and single-plant yield increased to varying degrees after the combined treatment of brassinolide and borage extract preparation, indicating that the borage extract preparation combined with the regulator treatment can promote the growth and fruit setting of tomato fruits, accelerate the color change of tomato fruits, and increase the yield of tomato fruits.

[0248] Table 22 Effect of Borago extract preparation treatment on tomato fruit growth

[0249]

[0250] Test Example 8

[0251] The effect of Borago extract preparation combined with potassium dihydrogen phosphate foliar spray on wheat yield and quality was tested.

[0252] 1. Test methods

[0253] (1) Test sample preparation

[0254] The Borago extract preparation prepared as described in Experimental Example 6, wherein the Borago extract was added in an amount of 20% (ie, the concentration of the effective substance was 200,000 ppm), was filter-sterilized and then diluted and applied according to the experimental design.

[0255] (2) Experimental design

[0256] Table 23 Experimental design

[0257]

[0258] 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.

[0259] (3) Experimental treatment

[0260] During the heading and filling stages of wheat, pesticides were sprayed according to the experimental design. When harvesting, the thousand-grain weight and plot yield of wheat were investigated, and samples were taken for quality testing.

[0261] 2. Results Analysis

[0262] Borage extract preparations applied at different growth stages of wheat can promote the increase of wheat thousand-grain weight and yield. Compared with the water control, the thousand-grain weight increased by 13.20% and the yield per square plot increased by 11.13%. When Borage extract preparations were applied in combination with potassium dihydrogen phosphate foliar fertilizer products, wheat yield was relatively improved, with the thousand-grain weight increasing from 44.6g to 45.7g and the yield per square plot increasing by 3.2g.

[0263] Table 24 Effect of Borage Extract Preparation on Wheat Yield

[0264]

[0265] Testing the quality of wheat grains treated with different methods showed that compared with pure water, the protein content and starch content in wheat grains treated with borage extract increased significantly, with growth rates reaching 16.55% and 15.41% respectively. The quality of wheat grains improved after borage extract preparation combined with potassium dihydrogen phosphate leaf spray treatment, with protein content increasing by 3.37% and starch content increasing by 3.88% compared with borage preparation alone.

[0266] 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.

[0267] Table 25 Effect of Borage Extract Preparation on Wheat Quality

[0268]

[0269] Test Example 9

[0270] To test the effect of leaf spraying of Borage extract preparation on increasing the yield and improving the quality of Chinese cabbage.

[0271] 1. Test methods

[0272] (1) Test sample preparation

[0273] In Experimental Example 6, the Borago extract preparation was prepared in an amount of 20% (ie, the concentration of the effective substance was 200,000 ppm). The preparation was sterilized by filtration and then diluted and applied according to the experimental design.

[0274] (2) Experimental design

[0275] Table 26 Experimental design

[0276]

[0277] (3) Experimental treatment

[0278] 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 leaf color and yield changes of the pakchoy were investigated at harvest, and samples were taken for determination of protein and vitamin C nutritional quality.

[0279] 2. Results Analysis

[0280] 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 to the water control treatment.

[0281] Table 27 Effect of leaf spraying of Borago extract preparation on the growth of Chinese cabbage

[0282]

[0283] After the leaves were sprayed with Borage extract, the vitamin C and protein contents of Chinese cabbage increased to varying degrees. Under the 250ppm concentration treatment, the quality improvement effect was better, with the vitamin C content increasing by 23.72% and the protein content increasing by 9.74%.

[0284] Table 28 Effect of leaf spraying of Borage extract preparation on the quality of Chinese cabbage

[0285]

[0286] Overall, leaf 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 part of Chinese cabbage and increase the content of vitamin C and protein.

[0287] Although 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 the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. Use of Borago officinalis extract in promoting plant growth and / or regulating plant stress resistance, characterized in that: The borage extract is an alcohol extract, the concentration of the borage extract is 0.001ppm-100ppm, the borage extract can promote seed germination and / or the growth of one or more of roots, stems, leaves or flowers and fruits, and the stress resistance is salt resistance and cold resistance.

2. The use according to claim 1, characterized in that: The alcohol solution is selected from methanol or ethanol.

3. The use according to claim 2, characterized in that: The alcohol solution is ethanol; the concentration of ethanol is 10% to 100%.

4. The use according to claim 3, characterized in that: The concentration of the ethanol is 40% to 80%.

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

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

20.

7. The use according to claim 1, characterized in that: The concentration of the borage extract is 2-50 ppm.

8. The use according to claim 1, characterized in that: The alcohol extract of Borage is applied to the following plants: Chinese cabbage, corn, rice, pepper, cucumber, tomato and wheat.

9. 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.

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

11. The application according to claim 10, 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.

12. The use according to claim 11, 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.

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