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CN-122011116-A - Salvia SmCLE-3 polypeptide and preparation method and application thereof

CN122011116ACN 122011116 ACN122011116 ACN 122011116ACN-122011116-A

Abstract

The invention relates to the field of preparation and application of biological peptides, in particular to a salvia SmCLE45-3 polypeptide, a preparation method and application thereof. The polypeptide amino acid sequence is shown as SEQ ID NO.1, and the invention can efficiently and synergistically activate the metabolic pathways of the terpenes and the phenylpropanes of the salvia miltiorrhiza by utilizing the systematic regulation and control function of the plant endogenous signal peptide, and synchronously and obviously promote the biomass of the root of the salvia miltiorrhiza and the content and accumulation of tanshinone IIA, cryptotanshinone and salvianolic acid B on the premise of not inhibiting the growth of the plants. The technology has the advantages of accurate action, safety, environmental protection, low dosage, simple and convenient operation and the like, and is suitable for high-quality and high-yield standardized cultivation of the red sage root.

Inventors

  • LIN LIHUA
  • Di Huanhuan

Assignees

  • 易博生物科技(青岛)有限公司

Dates

Publication Date
20260512
Application Date
20260204

Claims (10)

  1. 1. A Salvia miltiorrhiza SmCLE-3 polypeptide is characterized in that the amino acid sequence is shown as SEQ ID NO. 1.
  2. 2. The method for preparing the salvia miltiorrhiza SmCLE45-3 polypeptide of claim 1, which is characterized by being synthesized manually.
  3. 3. Use of the polypeptide SmCLE, 45-3 of red sage root in the preparation of a preparation for increasing active ingredient content of red sage root according to claim 1.
  4. 4. The method according to claim 3, wherein the active ingredients of the Salvia Miltiorrhiza are tanshinone IIA, cryptotanshinone and salvianolic acid B.
  5. 5. The method of claim 3, wherein the effective working concentration of the Salvia miltiorrhiza SmCLE45-3 polypeptide in the preparation is 0.5 nM-500 nM.
  6. 6. The method according to claim 3, wherein the solvent of the preparation is sterile deionized water or a buffer system comprising the auxiliary components of 0.1-1.0 mM calcium chloride, 0.005-0.02% by volume of wetting and penetrating agent of nonionic surfactant, and 2-10 mM MES or phosphate buffer to adjust the pH to 5.5-6.5.
  7. 7. A process for the preparation of a formulation as claimed in claim 3, comprising the steps of: (1) Synthesizing a salvia SmCLE-3 peptide segment shown in SEQ ID NO. 1 by adopting a solid phase synthesis method; (2) Freeze-drying the purified peptide fragment into powder, and drying and preserving at-20 ℃ or-80 ℃; (3) When in use, a proper amount of freeze-dried powder is taken and dissolved by a solvent to prepare a high-concentration stock solution; (4) The stock solution was diluted with the solvent to the desired working concentration prior to application, ready-to-use, or stored for a short period at 4 ℃.
  8. 8. A method for improving the content of active ingredients in the root of red-rooted salvia by using the preparation according to claim 3, which is characterized in that the preparation solution is uniformly sprayed on plant leaves in the vegetative growth phase and the root active ingredient accumulation phase of the root of the red-rooted salvia by a spraying mode.
  9. 9. The method of claim 8, comprising the steps of: (1) The first stage comprises spraying Saviae Miltiorrhizae radix plant after transplanting for 30-50 days when the plant has 4-6 true leaves; (2) And the second stage, spraying the red sage root in the early bud stage of 80-110 days after transplanting.
  10. 10. A salvia miltiorrhiza medicinal material cultivated by the method of claim 8 or 9.

Description

Salvia SmCLE-3 polypeptide and preparation method and application thereof Technical Field The invention relates to the field of preparation and application of biological peptides, in particular to a salvia SmCLE45-3 polypeptide, a preparation method and application thereof. Background The medicinal value of the red sage root as a traditional bulk medicinal material in China mainly depends on two major active ingredients, namely fat-soluble tanshinone (such as tanshinone IIA and cryptotanshinone) and water-soluble salvianolic acid (such as salvianolic acid B). The two components have definite pharmacological activity in the aspect of treating cardiovascular and cerebrovascular diseases, and are core indexes for evaluating the quality of the salvia miltiorrhiza medicinal materials. However, in actual production, the content of the active ingredients of the red sage root is affected by various factors such as genetic background, growth environment, cultivation period, harvesting time and the like, the fluctuation is large, the stability is poor, and the method becomes a key bottleneck for restricting the high-quality development of the red sage root industry. At present, strategies for improving the active ingredients of the red sage root mainly comprise genetic improvement, cultivation optimization and exogenous material regulation. Although exogenous application of plant hormone (such as methyl jasmonate and salicylic acid) or chemical inducer can stimulate secondary metabolism to a certain extent, the plant hormone has the problems of nonspecific action, narrow concentration window, possibility of inhibiting growth or bringing residual risk, and the like, and is difficult to apply in large scale in green and sustainable Chinese medicinal material production. The biosynthesis of tanshinone belongs to a plant terpenoid metabolic pathway, originates from Mevalonate (MVA) and methylerythritol phosphate (MEP) pathways, is synthesized by geranylgeranyl pyrophosphate (GGPP), and forms a tanshinone backbone under the catalysis of key enzymes such as katakiyl pyrophosphate synthase (CPS) and Kaurene Synthase (KSL). Salvianolic acid belongs to a phenylpropane metabolic pathway, and synthesis of the salvianolic acid is regulated and controlled by key enzymes such as Phenylalanine Ammonia Lyase (PAL), 4-coumaroyl-CoA ligase (4 CL) and the like. Both of these pathways are tightly regulated by complex transcriptional networks and signal pathways. In recent years, research on plant endogenous small peptide signal molecules provides a new idea for accurate regulation and control of crop traits. CLE (CLAVATA 3/EMBRYO SURROUNDING REGION-RELATED) family polypeptides are an important class of intercellular signal molecules and are involved in regulating and controlling various biological processes such as plant stem cell maintenance, organ development, nutrition distribution, stress response and the like. Some CLE peptides have been shown to act as long distance signals, systematically regulating root cap structure, nitrogen and phosphorus absorption and secondary metabolic flows. Chinese patent CN121293312A discloses a SmCLE mature peptide and application thereof in plant stress resistance, and the technical scheme comprises a SmCLE mature peptide, wherein the amino acid sequence of the SmCLE mature peptide is SEQ ID NO. 1. The invention is applied to the aspect of plant stress resistance, solves the problem that the existing CLE peptide can not be applied to stress resistance of plants, especially medicinal plants or non-mode crops, has high biocompatibility and targeting property, and can obviously improve biomass and yield of crops under stress environment. Chinese patent CN121227797A discloses application of SmCLE mature peptide in inhibiting plant senescence, and the amino acid sequence of SmCLE mature peptide is SEQ ID NO. 1. The invention is applied to the aspect of inhibiting plant senescence, solves the problem that the existing CLE peptide cannot be applied to the senescence resistance of plants, especially medicinal plants or non-mode crops, and has the characteristics of small molecular weight, easy synthesis, low action concentration and good biocompatibility, and can realize the precise regulation and control of leaf senescence by exogenous application, thereby avoiding the safety problem of transgenic organisms. However, whether SmCLE peptide has positive effect on improving plant active ingredients or not has not been reported yet, and whether the effect of promoting the biosynthesis of tanshinone/salvianolic acid components can be achieved by applying SmCLE peptide has yet to be studied. Disclosure of Invention The invention aims to solve the technical problems that the current red sage root cultivation has low and unstable active ingredient content and lacks an efficient green special regulation and control product, the current research on SmCLE peptide is mainly focused on improving plant stress resistance