An agricultural bio-stimulant formulation, its preparation method and application

Abstract

The application discloses an agricultural biological stimulant preparation and a preparation method and application thereof, and belongs to the field of agricultural biological stimulant preparation. The agricultural biological stimulant preparation comprises the following components (in percentage by weight): 5-50% of water extract of by-product of Auricularia auricular's fruiting body, 1-20% of thymol, 0.5-10% of Tween, 1-10% of ethyl acetate and 2-80% of water. The thymol in the agricultural biological stimulant preparation is sufficient in the market and easy to obtain. The water extract of the mycorrhiza and / or the fruiting body of Auricularia auricular is a resource recycling of by-product of agricultural products, and is safe to the environment. The agricultural biological stimulant preparation is suitable for green and organic agricultural product production, and has remarkable economic and social benefits and wide application prospect. The biological stimulant preparation is used in foliar spraying of green vegetables, so that the yield and quality of the green vegetables can be remarkably improved.

Description

TECHNICAL FIELD

[0001] The present application relates to an agricultural biological stimulant preparation and its preparation method and application, and belongs to the technical field of agricultural production. BACKGROUND

[0002] Green vegetable (scientific name: Brassica chinensis) is a large-scale vegetable crop with high consumption in China. Due to its high demand throughout the year, it is currently mainly cultivated in facilities. The high frequency of cultivation for many years has led to a decline in soil quality and serious nutrient loss in facility land, requiring continuous increase in chemical fertilizer input to maintain vegetable growth. However, this can cause continuous destruction of the agricultural environment and seriously affect the yield and quality of green vegetables. Therefore, under the premise of reducing the use of chemical inputs, how to improve the yield and quality of green vegetables is an important development direction to ensure the sustainable development of facility agriculture.

[0003] Using biological stimulants to regulate crop growth and quality is currently a research hotspot. Natural biological stimulants refer to natural compounds developed from natural resources, which have the characteristics of promoting crop growth, improving crop stress resistance, and being friendly to the environment. Using natural biological stimulants to improve vegetable yield and quality can reduce the use of chemical fertilizers, benefit the restoration and sustainable planting of facility vegetable production environment, and reduce resource waste. Currently, biological stimulants are mostly natural active ingredients from natural resources, with low extraction yield and high cost; they often have effects in the laboratory but unstable effects in the field. Therefore, developing a natural biological stimulant preparation with low cost, easy operation, and the ability to play a role in the field is an important measure to ensure the safe and sustainable production of facility vegetables.

[0004] Thymol is the main active ingredient of essential oil of Thymus L. plants in Lamiaceae, which is widely used in medicine and food preservation due to its bactericidal activity and environmental friendliness. Currently, thymol has been produced on a large scale, and compared with other natural compounds that can only be extracted, its source is more stable and suitable for large-scale production. As a natural active phenolic compound of plant origin, it is speculated that thymol may have the potential to regulate crop physiology, but as a natural phenolic compound, it may have unstable activity in the field. Therefore, it needs to be made into a suitable preparation for use in field agriculture. The by-product of Pleurotus cornucopiae refers to a large amount of inedible parts (irregularly shaped mushrooms, stipes, and mycorrhizae, etc.) generated during the picking and processing of Pleurotus cornucopiae, accounting for about 5% to 10% of the mass of fresh mushrooms. The current treatment method is to directly discard or burn the by-product of Pleurotus cornucopiae, causing resource waste and affecting the ecological environment. If the by-product of Pleurotus cornucopiae can be resourcefully utilized, it can be turned into treasure. The by-product of Pleurotus cornucopiae has a variety of active ingredients and has great potential to regulate crop growth and quality as an agricultural biological stimulant; other polysaccharide components in it may contribute to the stability of the agricultural preparation. Therefore, it is speculated that the mixed preparation of thymol and Pleurotus cornucopiae by-product extract may play the role of a biological stimulant in the field. SUMMARY

[0005] The first object of the present application is to provide an agricultural bio-stimulant, the second object of the present application is to provide a preparation method of the agricultural bio-stimulant, and the third object of the present application is to provide the application of the bio-stimulant in improving the growth and quality of green vegetables.

[0006] Technical solution: The agricultural bio-stimulant comprises the following components (by weight percentage): 5-50% of water extract of seafood mushroom by-product, 1-20% of thymol, 0.5-10% of Tween, 1-10% of ethyl acetate, and 2-80% of water.

[0007] Further, the agricultural bio-stimulant comprises the following components (by weight percentage): 20-40% of water extract of seafood mushroom by-product, 7-16% of thymol, 2-10% of Tween, 2-8% of ethyl acetate, and 2-80% of water.

[0008] Further, the seafood mushroom by-product comprises inedible seafood mushroom mycorrhizae and / or seafood mushroom stipe, and the Tween is Tween 60 or Tween 80.

[0009] Further, the preparation of the water extract of seafood mushroom by-product comprises the following steps:

[0010] The seafood mushroom by-product is washed with sterile water, dried, mixed with sterile water, crushed into a slurry, extracted in a water bath under stirring, cooled at room temperature, centrifuged, and the supernatant is obtained as the extract of seafood mushroom by-product.

[0011] Further, the volume ratio of the seafood mushroom by-product to sterile water is 1:1-3.

[0012] Further, the water bath extraction under stirring is carried out at 40-60°C for 3-5h, and the centrifugation is carried out at 8000-12000rpm for 30-60min.

[0013] The preparation method of the agricultural bio-stimulant comprises the following steps:

[0014] (1) uniformly mixing thymol, Tween, and ethyl acetate to form a solution S1;

[0015] (2) uniformly mixing the water extract of seafood mushroom by-product with water to form a solution S2;

[0016] (3) mixing the solution S1 with the solution S2, and high-speed shearing stirring with an emulsifying shearing machine to form a uniform emulsion.

[0017] Further, in step (3), the high-speed shearing stirring is at 1500-2500 rpm for 30-50 min.

[0018] The application also includes the use of the agricultural biological stimulator in improving the growth and quality of green vegetables.

[0019] Further, the agricultural biological stimulator is diluted with water to form a working solution, which is sprayed on the surface of the green vegetable leaves.

[0020] Further, the agricultural biological stimulator is sprayed once on the 30th and 45th day of growth, and the amount of the agricultural biological stimulator is sufficient to evenly wet the surface of the green vegetable leaves without droplet aggregation.

[0021] Advantages: Compared with the prior art, the present application has the following advantages:

[0022] (1) One of the main raw materials used in the agricultural biological stimulator is thymol, which is a natural food additive allowed by the state and has been widely used in the food industry. The market supply is sufficient and easy to obtain. The other main raw material is the water extract of seafood mushroom by-product, which is the mycorrhiza and / or stipe of seafood mushroom. It is a resource recycling of agricultural by-products and is safe to the environment.

[0023] (2) The agricultural biological stimulator is suitable for green and organic agricultural product production, and has significant economic and social benefits, with broad application prospects.

[0024] (3) The biological stimulator can significantly improve the yield and quality of green vegetables when used for foliar spraying. DETAILED DESCRIPTION

[0025] The technical solutions of the present application are further described below.

[0026] Example 1

[0027] 1. Preparation of water extract of seafood mushroom by-product:

[0028] Fresh seafood mushroom by-products (non-edible mycorrhiza and stipe) were washed with sterile water and dried, then mixed with sterile water at a weight ratio of 1:1, and broken into slurry with a high-speed multifunctional grinder. The slurry was extracted at 50℃ for 3h, and stirred at a constant speed during the extraction. After cooling at room temperature, centrifugation was performed at 12000 rpm for 30 min, and the supernatant, i.e. the water extract of seafood mushroom by-product, was prepared for use.

[0029] 2. Preparation of agricultural biological stimulator:

[0030] According to the percentage by weight: 10% of the water extract of seafood mushroom by-product, 4% of thymol, 2% of Tween 80, 5% of ethyl acetate, and 79% of water.

[0031] (1) Thymol, Tween, and ethyl acetate were mixed uniformly at room temperature to form solution S1.

[0032] (2) The water extract of seafood mushroom by-product and water were mixed uniformly to form solution S2.

[0033] (3) Solutions S1 and S2 were mixed, and an emulsifying shear machine was used to shear and stir at 2500 rpm for 40-50 min to form a uniform emulsion, thereby obtaining the agricultural biological stimulant preparation.

[0034] (4) The prepared agricultural biological stimulant preparation was diluted 5 times with water to obtain working solution S3, which was used for field spraying.

[0035] Comparative Example 1

[0036] The preparation process was the same as in Example 1, except that 14% of thymol, 2% of Tween 80, 5% of ethyl acetate, and 79% of water were weighed according to the percentage by weight to prepare working solution S4.

[0037] Comparative Example 2

[0038] The preparation process was the same as in Example 1, except that 14% of the water extract of seafood mushroom by-product, 2% of Tween 80, 5% of ethyl acetate, and 79% of water were weighed according to the percentage by weight to prepare working solution S5.

[0039] Example 2 Field Spraying of Agricultural Biological Stimulant Preparation

[0040] Greenhouse-grown green vegetables were selected, and uniform growth was selected for the field plot, with 4 groups of treatments. Treatment 1 was sprayed with water as a control, treatment 2 was sprayed with working solution S3 in Example 1. Treatment 3 was sprayed with working solution S4 in Comparative Example 1, and treatment 4 was sprayed with working solution S5 in Comparative Example 2. Each treatment set up 3 small areas as repeats, with an area of 20 m 2 , arranged randomly in groups.

[0041] The growth period of green vegetables in the greenhouse in this experiment was 50 days, and each was sprayed once on the 30th and 45th day. The spraying amount was sufficient to evenly wet the leaf surface without liquid droplets coagulating and falling. In addition to the above spraying treatment, the green vegetables in each treatment group were managed according to the local conventional cultivation management measures. After the green vegetables were harvested, five-point sampling method was used to take green vegetable samples from each treatment group, and the biomass and various nutritional quality indicators were measured. Among them, fresh weight and dry weight were measured by weighing method, and leaf thickness was measured by measuring method, as shown in Table 1.

[0042] Table 1 Effects of spraying the biological stimulator on the growth of Chinese cabbage

[0043]

[0044] Note: * indicates that the treatment and the control are significantly different at the level of P < 0.05.

[0045] As can be seen from Table 1, after spraying the agricultural biological stimulator prepared in Example 1, the fresh weight, dry weight, water content and leaf thickness of Chinese cabbage were increased by 32.85%, 24.31%, 2.27% and 45.21% respectively compared with the water control, while the growth-promoting effect of thymol preparation (S4) or water extract of Hypsizygus mabroscus by-product preparation (S5) alone was not significant. It is shown that spraying the biological stimulator of the present application can significantly promote the growth of Chinese cabbage.

[0046] The chlorophyll content determination method refers to NY / T 3082-2017, and the carotenoid content determination method refers to GB5009.83-2016. The results are shown in Table 2.

[0047] Table 2 Effects of spraying the biological stimulator on the chlorophyll content of Chinese cabbage leaves

[0048]

[0049] Note: * indicates that the treatment and the control are significantly different at the level of P < 0.05.

[0050] As can be seen from Table 2, after spraying the agricultural biological stimulator prepared in Example 1, the contents of chlorophyll a, chlorophyll b and carotenoids in Chinese cabbage were higher than those of thymol preparation (S4) or water extract of Hypsizygus mabroscus by-product preparation (S5) alone and the water control. It is shown that spraying the biological stimulator of the present application can significantly promote the growth of Chinese cabbage, and this growth-promoting effect may be due to the significant increase in total chlorophyll content, thereby improving photosynthetic performance.

[0051] The vitamin C content determination method refers to GB / T 6195-1986, the soluble protein content determination method adopts the Coomassie brilliant blue method, and the soluble sugar content determination method refers to GB / T 36056-2018. The results are shown in Table 3.

[0052] Table 3 Effects of spraying the biological stimulator on the soluble protein, vitamin C and soluble sugar contents of Chinese cabbage leaves

[0053] Treatment Vitamin C (mg / kg) Soluble protein (mg / g) Soluble sugar (mg / g) Treatment 1 (control) 135.20±9.13 6.84±0.37 12.31±1.98 Treatment 2 (S3) 201.32±12.56* 7.12±0.12* 12.95±1.35 Treatment 3 (S4) 141±5.57 6.85±0.02 12.64±0.94 Treatment 4 (S5) 138±6.89 6.92±0.04 12.23±0.87

[0054] Note: * indicates that the treatment and the control are significantly different at the level of P < 0.05.

[0055] As can be seen from Table 3, the vitamin C content in the green vegetables after spraying the agricultural biological stimulant preparation prepared in Example 1 was significantly increased by 48.91% compared with the control; the soluble protein content was increased by 4.09% compared with the control; the soluble sugar had no significant change; and the improvement effect was better than that of thymol preparation (S4) or water extract of seafood mushroom by-product preparation (S5) alone. It is indicated that spraying the biological stimulant preparation of the application can significantly improve the quality of green vegetables.

[0056] The free amino acid content determination method refers to GB / T 30987-2020, and the results are shown in Table 4.

[0057] Table 4 Effect of spraying biological stimulant on the amino acid content of green vegetable leaves

[0058] Amino acids (g / 100 g) Treatment 1 (control) Treatment 2 (S3) Treatment 3 (S4) Treatment 4 (S5) Proline 0.016 0.034 0.024 0.024 Glycine 0.020 0.038 0.032 0.028 Alanine 0.031 0.053 0.050 0.041 Valine 0.027 0.044 0.035 0.037 Methionine 0.0087 0.014 0.011 0 Isoleucine 0.015 0.030 0.029 0.020 Aspartic acid 0.035 0.073 0.061 0.047 Threonine 0.016 0.034 0.029 0.021 Serine 0.023 0.041 0.039 0.029 Glutamic acid 0.15 0.230 0.230 0.20 Leucine 0.025 0.054 0.049 0.035 Phenylalanine 0.013 0.035 0.027 0.020 Histidine 0.0089 0.019 0.017 0.011 Lysine 0.026 0.053 0.047 0.034 Arginine 0.012 0.033 0.028 0.019 Cystine 0 0.012 0.011 0

[0059] As can be seen from Table 4, the various amino acid contents in the green vegetables after spraying the agricultural biological stimulant preparation prepared in Example 1 were also significantly increased by 53.3%-175.0% compared with the control, and the improvement effect was better than that of thymol preparation (S4) or water extract of seafood mushroom by-product preparation (S5) alone. It is indicated that spraying the biological stimulant preparation of the application can significantly improve the quality of green vegetables.

[0060] Example 3

[0061] 1. Preparation of water extract of seafood mushroom by-product, same as Example 1.

[0062] 2. Preparation of agricultural biological stimulant preparation:

[0063] According to the weight percentage, take: treatment 5: water extract of seafood mushroom by-product 51%, thymol 21%, Tween 80 4%, ethyl acetate 5%, water 19%; treatment 6: water extract of seafood mushroom by-product 5%, thymol 20%, Tween 4%, ethyl acetate 5%, water 46%; treatment 7: water extract of seafood mushroom by-product 50%, thymol 1%, Tween 4%, ethyl acetate 5%, water 40%, preparation process same as Example 1. The working solution prepared is S6, S7, S8 respectively.

[0064] In this experiment, the growth period of the facility greenhouse green vegetables was 50 days, and each was sprayed once on the 30th day and the 45th day. The spraying amount was sufficient and uniform wetting of the leaf surface without liquid droplet condensation and falling. In addition to the above spraying treatment, the green vegetables of the two groups were managed according to the local conventional cultivation management measures. After the green vegetables were harvested, five-point sampling method was used to take green vegetable samples of the two groups, and the biomass was determined. Among them, the fresh weight and dry weight were determined by weighing method, and the leaf thickness was determined by measuring method, and the results are shown in Table 5.

[0065] Table 5 Effects of spraying the biological stimulant on the growth of Chinese flowering cabbage

[0066]

[0067] As shown in Table 5, the fresh weight, dry weight, water content and leaf thickness of the Chinese flowering cabbage are most obviously increased after spraying the biological stimulant preparation of Example 1. When the ratio of thymol and the extract of the by-product of the Hypsizygus marmoreus in the biological stimulant preparation is more than the concentration range specified in the present application, the preparation formed does not obviously promote the growth of the Chinese flowering cabbage.

Claims

1. An agricultural biostimulant preparation, characterized in that, The agricultural biostimulant preparation comprises the following components by weight percentage: 5%–50% water extract of seafood mushroom by-products, 1%–20% thymol, 0.5%–10% Tween, 1%–10% ethyl acetate, and 2%–80% water. The seafood mushroom by-products include inedible seafood mushroom mycorrhizae and / or seafood mushroom stipes. The preparation of the aqueous extract of the seafood mushroom by-product includes the following steps: Wash the seafood mushroom by-products with sterile water, drain the water, add sterile water and mix. The volume ratio of seafood mushroom by-products to sterile water is 1:1~3. Crush into a pulp, extract the pulp in a water bath at 40-60℃ for 3-5 minutes, cool at room temperature, centrifuge, and take the supernatant, which is the water extract of seafood mushroom by-products.

2. The agricultural biostimulant preparation according to claim 1, characterized in that, The agricultural biostimulant preparation comprises the following components by weight percentage: 20%~40% water extract of seafood mushroom by-product, 7%~16% thymol, 2%~10% Tween, 2%~8% ethyl acetate, and 2%~80% water.

3. The agricultural biostimulant preparation according to claim 1, characterized in that, The Tween is either Tween 60 or Tween 80.

4. The method for preparing the agricultural biostimulant preparation according to any one of claims 1-3, characterized in that, Includes the following steps: (1) Mix thymol, Tween and ethyl acetate evenly to form solution S1; (2) Mix the aqueous extract of the seafood mushroom by-product with water to form solution S2; (3) Mix solution S1 and solution S2, and use an emulsifying shearing machine to shear and stir at high speed to form a homogeneous emulsion liquid.

5. The preparation method according to claim 4, characterized in that, The high-speed shearing and stirring is carried out at 1500-2500 rpm for 30-50 minutes.

6. The application of the agricultural biostimulant preparation according to any one of claims 1-3 in improving the growth and quality of leafy greens.

7. The application according to claim 6, characterized in that, The agricultural biostimulant preparation described in any one of claims 1-3 is diluted with water to form a working solution, and the working solution is sprayed onto the surface of the leafy greens.

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