CN-122012645-A - Preparation method of leupeptin and application of leupeptin in agriculture

Abstract

The invention relates to the field of agricultural biology, and particularly discloses a preparation method of a leupeptin and application thereof in agriculture, wherein the preparation method comprises the following steps of S1, carrying out liquid fermentation on a tray Mi Qingmei (Penicillium thomii) with a preservation number of CGMCC No.42204 to obtain mycelium, S2, carrying out ultrasonic auxiliary extraction on the mycelium by adopting an ethanol solution to obtain a crude extract, S3, carrying out chemical synthesis modification on precursor substances in the crude extract, including Boc protection linear dipeptide synthesis, protecting group removal and cyclization reaction, to obtain the leupeptin, and constructing a synergistic composition application system by scientifically compounding the leupeptin derived from a specific Tomicis strain, alginic acid and sodium nitrate.

Inventors

• JIAO YUZHU
• WANG BIN
• GUO ZIBO
• LIU CHANGQING
• SUN JIAN
• WANG HONGFENG
• DAI LILI

Assignees

• 山东蓬勃生物科技有限公司

Dates

Publication Date

20260512

Application Date

20251122

Claims (7)

1. 1. A method for preparing a leupeptin, comprising the steps of: S1, carrying out liquid fermentation on penicillium tolmi with the preservation number of CGMCC NO.42204 to obtain mycelium; s2, carrying out ultrasonic auxiliary extraction on the mycelium by adopting an ethanol solution to obtain a crude extract; s3, carrying out chemical synthesis modification on precursor substances in the crude extract, wherein the chemical synthesis modification comprises synthesis of Boc protection linear dipeptide, removal of a protecting group and cyclization reaction, so as to obtain the British peptide.
2. 2. The method of claim 1, wherein the leupeptin is mixed with agriculturally acceptable adjuvants when used in the preparation of stress-resistant compositions.
3. 3. The preparation method of the leupeptin according to claim 2, wherein alginic acid and sodium nitrate are further added in the preparation of the stress-resistant composition, wherein the effective concentration of the leupeptin in the final composition is 5 ng/mL-100 ng/mL, the effective concentration of alginic acid is 10 mg/L-100 mg/L, and the effective concentration of sodium nitrate is 1 mg/L-10 mg/L.
4. 4. A method of preparing a leupeptin according to claim 3, wherein the effective concentration of said leupeptin in the final composition is 20ng/mL, the effective concentration of said alginic acid is 40mg/L, and the effective concentration of said sodium complex nitrate is 3mg/L.
5. 5. Use of the leucins prepared according to the method of claim 1 in agriculture, wherein the leucins are applied to plants to relieve adversity stress suffered by the plants, wherein the adversity stress is salt stress, low temperature stress or drought stress.
6. 6. The use according to claim 5, wherein the plant is maize, lettuce or cucumber.
7. 7. The use according to claim 5 or 6, wherein said alleviation of plant stress is manifested as one or more of the following effects: improving the survival rate, fresh weight, dry weight or root length of plants; Maintaining a higher chlorophyll content; enhancing the activity of superoxide dismutase, peroxidase or catalase; reducing malondialdehyde accumulation; The higher K + /Na + ratio in the cell was maintained.

Description

Preparation method of leupeptin and application of leupeptin in agriculture Technical Field The invention belongs to the field of agricultural biology, and particularly relates to a preparation method of a Bright peptide and application thereof in agriculture. Background Plants are often subjected to a variety of abiotic stresses throughout the growth cycle, with soil salinization, low temperature and drought being the primary environmental factors that limit crop yield and quality improvement. These stress conditions can cause a series of physiological disturbances such as imbalance in the osmotic regulation of plants, disruption of ion homeostasis, accumulation of active oxygen in large amounts, and damage to cell membrane structures, ultimately leading to plant growth inhibition, biomass decline, and yield loss. In order to solve the problems, measures such as physical improvement, stress-resistant variety breeding or application of exogenous plant growth regulators are commonly adopted in agricultural production. Among them, administration of exogenous regulators has received attention because of easy operation and quick effect. Various biological or chemical agents have been used to enhance stress resistance of plants, such as alginic acid, sodium nitrite and the like, which have a certain biostimulation or growth regulation function. However, most products in the prior art still have the limitations of single function, unstable action and effect, higher effective application concentration, poor universality among different crops and stress types and the like, and the comprehensive requirements of modern agriculture on green, efficient and multifunctional stress-resistant technologies are difficult to meet. Disclosure of Invention In order to solve the technical problems, the invention provides application of a composition based on a penicillium tolmi extract in alleviating plant stress, so as to solve the problem that a plant stress-resistant technical scheme with broad-spectrum stress resistance, high-efficiency synergistic effect and environmental friendliness at an ultralow concentration is lacking in the prior art. A method for preparing a leupeptin, comprising the steps of: S1, carrying out liquid fermentation on a tray Mi Qingmei (Penicillium thomii) with the preservation number of CGMCC NO.42204 to obtain mycelium; s2, carrying out ultrasonic auxiliary extraction on the mycelium by adopting an ethanol solution to obtain a crude extract; s3, carrying out chemical synthesis modification on precursor substances in the crude extract, wherein the chemical synthesis modification comprises synthesis of Boc protection linear dipeptide, removal of a protecting group and cyclization reaction, so as to obtain the British peptide. Preferably, the leupeptin is mixed with agriculturally acceptable adjuvants when used for preparing stress-resistant compositions. Preferably, alginic acid and sodium nitrate are also added when the stress-resistant composition is prepared, the effective concentration of the British peptide in the final composition is 5 ng/mL-100 ng/mL, the effective concentration of the alginic acid is 10 mg/L-100 mg/L, and the effective concentration of the sodium nitrate is 1 mg/L-10 mg/L. Preferably, the effective concentration of the leupeptin in the final composition is 20ng/mL, the effective concentration of alginic acid is 40mg/L, and the effective concentration of sodium complex nitrate is 3mg/L. The application of the leucins prepared by the method in agriculture applies the leucins to plants to relieve the adversity stress suffered by the plants, wherein the adversity stress is salt stress, low temperature stress or drought stress. Preferably, the plant is maize, lettuce or cucumber. Preferably, said alleviation of plant stress is manifested as one or more of the following effects: improving the survival rate, fresh weight, dry weight or root length of plants; Maintaining a higher chlorophyll content; enhancing the activity of superoxide dismutase (SOD), peroxidase (POD) or Catalase (CAT); Reducing Malondialdehyde (MDA) accumulation; The higher K+/Na+ ratio in the cell is maintained. The biological material preservation information of the invention is as follows: The strain PFJ-4 used in the invention is identified as penicillium tolmi (Penicillium thomii) which is preserved in China general microbiological culture Collection center (CGMCC) for 09 and 16 days in 2025, and has the address of the institute of microorganisms, national academy of sciences of China, no. 3, north Star West Lu 1, kyowa, beijing, and the postal code of 100101. The preservation number is CGMCC No. 42204. The strain was in a viable state as measured by the collection at 2025, month 09 and 16. Compared with the prior art, the invention has the following beneficial effects: The application system of the synergistic composition is constructed by scientifically compounding the white light peptide derived from the sp