Salvia miltiorrhiza SmIDL1 polypeptide as well as preparation method and application thereof

Abstract

The invention relates to the field of preparation and application of biological peptides, in particular to a salvia miltiorrhiza SmIDL1 polypeptide as well as a preparation method and application thereof. The amino acid sequence of the polypeptide is as shown in SEQ ID NO. 1; according to the method disclosed by the invention, by simulating an endogenous senescence / nutrition re-mobilization signal, the directional transport of a photosynthetic product to the root and the preferential distribution of the photosynthetic product to tanshinone biosynthesis are accurately regulated and controlled, so that the specific efficient accumulation of tanshinone components is realized on the premise of not remarkably promoting vegetative growth. The invention provides a brand new precise agricultural strategy of material flow regulation and quality improvement, and has important value for producing high-quality salvia miltiorrhiza medicinal materials with high tanshinone content.

Description

A Tanshinone SmIDL1 polypeptide, its preparation method and application Technical Field

[0001] This invention relates to the field of preparation and application of bioactive peptides, and particularly to a Danshen SmIDL1 polypeptide, its preparation method, and its application. Background Technology

[0002] The medicinal value of Salvia miltiorrhiza depends on two main categories of components: tanshinones (lipid-soluble) and salvianolic acids (water-soluble). Among them, tanshinone IIA and cryptotanshinone play a core role in the treatment of cardiovascular and cerebrovascular diseases. However, their content in the plant is usually low and fluctuates greatly due to environmental influences, which is a key bottleneck in improving the quality of Salvia miltiorrhiza.

[0003] Current methods for increasing tanshinone content, such as exogenous application of hormones like methyl jasmonate (MeJA), are effective but often accompanied by growth inhibition or non-specific effects. In recent years, endogenous plant peptide hormones have become a research hotspot due to their high activity, high specificity, and low residue. IDA / IDL (Influence Deficient in Abscision / IDA-Like) family peptides are key signaling molecules regulating floral organ abscission, leaf senescence, and lateral root development. They trigger downstream signals by binding to cell membrane receptor kinases, finely regulating cell separation and nutrient remobilization processes. For example, Chinese patent CN112400881A discloses a mature IDL7 peptide plant senescence promoter, its preparation method, and its application. This plant senescence promoter can accelerate leaf senescence without other adverse effects and has strong field operability. Chinese patent CN106614586A discloses a mature IDL4 peptide plant senescence promoter, its preparation method, and its application, which can accelerate leaf senescence without other adverse phenotypes. There are no reports, either domestically or internationally, of developing exogenous formulations of peptides associated with such "senescence-abscission" signals to actively regulate nutrient transport and targeted accumulation of secondary metabolites in medicinal plants. Summary of the Invention

[0004] The technical problem this invention aims to solve is the difficulty in increasing the content of tanshinone components in current Salvia miltiorrhiza production, which is often not coordinated with biomass growth. There are no reports, either domestically or internationally, on developing exogenous preparations of peptides related to the "senescence-abscission" signal for actively regulating nutrient transport and the targeted accumulation of secondary metabolites in medicinal plants. Whether the application of IDL peptides can increase the content of tanshinone IIA and cryptotanshinone in Salvia miltiorrhiza roots also requires further investigation.

[0005] To address the aforementioned issues, this invention provides a biological agent based on the novel signal peptide SmIDL1 and its application method. This agent can specifically and efficiently increase the content of tanshinone IIA and cryptotanshinone in the roots of *Salvia miltiorrhiza*, while having a limited or controllable promoting effect on the aboveground growth of the plant, thus achieving precise improvement with a "quality-first" approach. Furthermore, by controlling the spraying concentration and timing, the "source-sink" regulation principle is cleverly utilized in the later stages of *Salvia miltiorrhiza* growth to maximize the conversion efficiency of photosynthetic products into tanshinone biosynthesis, establishing a new cultivation and management scheme for *Salvia miltiorrhiza* with quality improvement as its core objective.

[0006] To achieve the above objectives, the present invention is implemented through the following technical means: a Danshen SmIDL1 polypeptide, the amino acid sequence of which is shown in SEQ ID NO.1.

[0007] SEQ ID NO. 1: IGQLPKGVPIPPSGPSNRTS.

[0008] Our team identified a new IDL family member in *Salvia miltiorrhiza*, named SmIDL1. It encodes a precursor protein that, after processing, produces a mature secretory peptide of 20 amino acids (sequence: IGQLPKGVPIPPSGPSNRTS). Preliminary expression analysis showed that the SmIDL1 gene was upregulated in senescent leaves and soon-to-be-abscissed organs of *Salvia miltiorrhiza*, suggesting its potential involvement in senescence signaling or nutrient redistribution regulation. However, no reports, either domestically or internationally, have documented the development of exogenous formulations of peptides associated with the "senescence-abscissence" signaling pathway for the active regulation of nutrient transport and the targeted accumulation of secondary metabolites in medicinal plants.

[0009] Tanshinone biosynthesis consumes a large amount of carbon skeleton and reducing power. Traditional yield-increasing methods (such as growth regulators) may not effectively increase tanshinone content due to the "dilution effect" or carbon flow competition, while increasing biomass. Therefore, a strategy is needed that can "remobilize" vegetative growth and even moderately guide vegetative substances to storage organs without significantly stimulating vegetative growth. Based on the core function of IDL peptides in regulating organ abscission and senescence-related nutrient cycling, we propose a novel strategy: exogenous application of SmIDL1 peptides to simulate endogenous senescence / remobilization signals. Without causing abnormal abscission, this strategy gently induces metabolic reprogramming at the leaf level, similar to early senescence, promoting the efficient transport of photosynthetic products (especially substrates flowing to terpene synthesis) to the roots and prioritizing their use in tanshinone biosynthesis.

[0010] Compared with existing technologies, the fundamental difference in the regulatory logic of this invention is that it does not indirectly enhance the composition by promoting overall growth or activating a broad-spectrum defense response. Instead, it achieves precise intervention in the "directional transport" and "targeted unloading" of carbon flows by simulating a specific physiological signal related to nutrient redistribution, thereby more directly and efficiently enhancing the accumulation of target products. This provides a completely new technical approach and tool for improving the quality of medicinal plants.

[0011] A method for preparing the above-mentioned SmIDL1 polypeptide from Tanshinone, wherein the polypeptide is synthesized artificially. A solid-phase synthesis method, such as the Fmoc method, is preferred.

[0012] The above-mentioned SmIDL1 polypeptide from Danshen is used in the preparation of formulations that increase the content of fat-soluble active ingredients in Danshen.

[0013] Furthermore, the active ingredients of tanshinone are tanshinone IIA and cryptotanshinone.

[0014] Furthermore, the effective working concentration of the Tanshinone SmIDL1 polypeptide in the formulation is 1.0 nM to 200 nM. Preferably, it is 10 nM to 50 nM; the most preferred concentration is 20 nM.

[0015] Furthermore, the solvent for the formulation is a weakly acidic buffer solution with a pH of 5.8-6.2, such as 5-10 mM MES. This environment is conducive to peptide stability and leaf absorption.

[0016] Furthermore, the formulation includes an energy metabolism synergist: 0.05-0.2 mM adenosine-5'-phosphate (AMP), designed to slightly perturb cellular energy state and synergistically enhance the SmIDL1-induced carbon flow redistribution effect; and a wetting agent: 0.005%-0.01% (v / v) of a plant-derived surfactant (such as tea saponin).

[0017] A method for preparing the above-mentioned formulation includes the following steps: solid-phase synthesis of the peptide SEQ ID NO: 1, HPLC purification (≥95%) followed by lyophilization; and dissolving it in the above-mentioned buffer solution to prepare a working solution for use, which should be prepared and used immediately.

[0018] A method for enhancing the content of active ingredients in *Salvia miltiorrhiza* using the aforementioned formulation involves spraying the formulation solution onto the mature leaves of the lower and middle parts of the plant during the early stage of reproductive growth initiation to before full bloom. This precisely regulates the source-sink relationship and secondary metabolic flow in *Salvia miltiorrhiza*, efficiently driving the conversion and accumulation of tanshinone compounds in the roots from photosynthetic products, making it suitable for the quality-oriented production of *Salvia miltiorrhiza*. The critical spraying window is 100 to 130 days after transplanting, i.e., from the early stage of reproductive growth initiation to before full bloom; at this time, the plant has established a sufficient biomass base, and spraying SmIDL1 can primarily affect the later allocation of photosynthetic products, rather than promoting vegetative growth.

[0019] Furthermore, the specific steps include: spraying an effective working concentration (e.g., 20 nM) of SmIDL1 formulation 110-115 days after transplanting.

[0020] Furthermore, the specific steps include: the first spraying is applied during the budding stage after transplanting; the second spraying is applied 10-15 days after the first spraying, i.e. before the full bloom stage, with an effective working concentration (e.g., 20 nM) of SmIDL1 formulation.

[0021] Furthermore, when spraying, focus on the underside of the mature functional leaves in the lower part of the plant to promote the absorption of signal peptides and initiate local aging-related metabolic transformations.

[0022] Furthermore, maintain a relative soil moisture content of 60-70% for one week before and after spraying to avoid drought or waterlogging interfering with the signal. Ensure sufficient phosphorus and potassium nutrition to support the energy needs of root metabolism. Combined with gentle, light bud removal (removing about 1 / 3 of the flower buds), this further reduces competition for assimilates from the reproductive pool, allowing the guiding effect of SmIDL1 to be more focused on the synthesis of medicinal components in the roots.

[0023] A type of Salvia miltiorrhiza medicinal material cultivated using the above method has significantly higher contents of tanshinone IIA and cryptotanshinone in its roots than conventionally cultivated Salvia miltiorrhiza without the use of this preparation, and the increase in tanshinone content is greater than the increase in root biomass.

[0024] The beneficial effects of this invention are as follows: For the first time, the biological functions of IDL family peptides in regulating organ aging and nutrient remobilization are utilized, transforming them into a "directed transport inducer." SmIDL1 treatment does not aim to strongly promote growth, but rather intelligently "reprograms" the metabolic output of the source leaves, guiding carbon flow to preferentially meet the needs of tanshinone synthesis in the roots, achieving a strategic shift from "promoting production" to "regulating logistics." This formulation has a highly significant and specific effect on increasing tanshinone IIA and cryptotanshinone levels. Under optimal conditions, the increase in tanshinone content can be significantly higher than the increase in root biomass, effectively solving the industry problem of "increasing yield without improving quality." After spraying, the plants show a slow and moderate decrease in chlorophyll content in the middle and lower leaves (a sign of metabolic activity conversion), but no premature aging or abnormal leaf drop. Aboveground growth is stable, root metabolism is active, achieving a new balance between "source" and "sink" at a higher metabolic level, resulting in overall plant health. The ingredients are natural and degrade quickly. The strategy of precise application in the later stages of growth results in a low total dosage and low cost. This invention is particularly suitable for use in production bases that pursue high-quality, high-value-added Danshen medicinal materials, such as targeted planting for the production of tanshinone extract raw materials. Unlike growth regulators that promote overall biomass or inducers that broadly activate defense responses (such as MeJA), this invention provides a specialized tool focused on optimizing the allocation of assimilates in the later stages of growth. It can be combined with early growth-promoting management measures to form a relay-style cultivation program that "ensures yield in the early stages and improves quality in the later stages." Attached Figure Description

[0025] Figure 1: Comparison of root status of different treatments after harvest of Salvia miltiorrhiza after spraying with SmIDL1 foliar agent and control.

[0026] Figure 2: Expression patterns of the SmIDL1 gene in different tissues and developmental stages of *Salvia miltiorrhiza*. A is a bar chart showing changes in root dry weight; B is a bar chart showing changes in the content of tanshinone IIA and cryptotanshinone; * indicates significant differences; ** indicates highly significant differences.

[0027] Figure 3: Effects of different concentrations of SmIDL1 on the growth and component accumulation of Salvia miltiorrhiza. * indicates significant difference; ** indicates highly significant difference.

[0028] Figure 4: Comparison of the effects of SmIDL1 treatment and methyl jasmonate (MeJA) treatment on Salvia miltiorrhiza. * indicates significant difference; ** indicates highly significant difference.

[0029] Figure 5: Effects of SmIDL1 treatment on the expression of key genes in leaves and roots. Detailed Implementation

[0030] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0031] Furthermore, unless otherwise specified, all methods used in the embodiments of this invention are common processing methods; and all materials used, unless otherwise specified, are obtained by purchasing from the market.

[0032] Example 1: Synthesis and formulation of SmIDL1 peptide.

[0033] The SmIDL1 peptide with the sequence IGQLPKGVPIPPSGPSNRTS was prepared by solid-phase synthesis, purified by HPLC (purity 97%), and lyophilized. Working solution preparation: The lyophilized powder was dissolved in 5 mM MES buffer (pH 6.0, containing 0.01% tea saponin) to prepare a 100 µM stock solution. Before use, it was diluted to a working concentration of 20 nM with the same buffer.

[0034] Example 2: Pot experiment to screen the optimal concentration and verify the specificity of quality improvement.

[0035] Two-year-old potted Salvia miltiorrhiza seedlings were used as the material. Five treatments were set up: CK (buffered saline) and SmIDL1 (5, 20, 50, 100 nM). The treatment was applied as a single foliar spray at the budding stage (110 days after transplanting). The results were measured after harvest.

[0036] The results are as follows:

[0037] As shown in Figure 1, the SmIDL1 polypeptide foliar spray at 20 nM had the best effect on promoting the growth of Salvia miltiorrhiza roots. As shown in Figure 3A, the 20 nM treatment had no significant effect on the aboveground growth, but it could moderately increase the root dry weight. As shown in Figure 2B, the 20 nM treatment increased the content of both by a much larger percentage than it increased the root dry weight, highlighting its "quality-improving specificity".

[0038] The 20 nM treatment moderately increased the root dry weight (+8.2%) while significantly increasing the content of tanshinone IIA (+50%) and cryptotanshinone (+66.7%), while having no significant effect on salvianolic acid B, confirming its quality improvement specificity and optimal concentration effect.

[0039] Example 3: To clarify the expression pattern of the SmIDL1 gene during the senescence process of *Salvia miltiorrhiza* leaves, we cultivated *Salvia miltiorrhiza* plants under strictly controlled light, temperature, and moisture conditions. Based on leaf morphology and physiological state, leaf samples were collected from four developmental stages: S1 (young leaves, fully expanded but not fully extended), S2 (extended leaves, fully expanded and dark green), S3 (pre-senescence leaves, showing the earliest visible senescence characteristics such as slight yellowing at the leaf tip), and S4 (mid-to-late senescence leaves, with significantly increased yellowing or wilting area). Root tissue was also collected as a control. At least three biological replicates were established for each stage. Samples were immediately flash-frozen in liquid nitrogen after collection and stored at -80°C. Total RNA was extracted from each sample in the laboratory using a total RNA extraction kit. After extraction, RNA concentration and purity were determined using a micro spectrophotometer (A260 / A280 ratio between 1.8 and 2.0), and RNA integrity was checked by 1% agarose gel electrophoresis to ensure clear 28S and 18S ribosomal RNA bands without significant degradation. One μg of high-purity total RNA was used to synthesize first-strand cDNA using a reverse transcription kit containing a gDNA removal step. The product was diluted and stored at -20°C for later use. Specific primers for the target gene SmIDL1 were designed, and primer specificity and amplification efficiency were verified by conventional PCR and melting curve analysis. Simultaneously, the housekeeping gene SmActin, which has been verified to be stably expressed in *Salvia miltiorrhiza*, was selected as an internal control. qRT-PCR reactions were performed using the SYBR Green dye method on a real-time quantitative PCR instrument. Three technical replicates were set up for each cDNA sample, and the reaction system was prepared strictly according to the premix instructions, with a template-free control included. The PCR procedure included: 95°C pre-denaturation, followed by 40 cycles of 95°C denaturation, 60°C annealing / extension, and fluorescence signal acquisition. Finally, melting curve analysis was performed to confirm the uniformity of the amplified products. Experimental data were compared using the Ct method. -ΔΔCt Relative quantitative analysis was performed. The S1 period samples with relatively stable expression were used as the standard group (with their relative expression level set to 1), and the relative expression levels of SmIDL1 at different periods were finally obtained.

[0040] As shown in Figure 2, qRT-PCR analysis revealed that SmIDL1 expression was highest in leaves during the pre-senescence stage, and lower in young tissues and roots, suggesting its association with the senescence process. S1: young leaves; S2: spreading leaves; S3: leaves showing senescence characteristics, i.e., leaves in the pre-senescence stage; S4: leaves in the middle and late senescence stages.

[0041] Example 4: Healthy and uniformly growing *Salvia miltiorrhiza* seedlings were selected and transplanted into uniformly sized planting pots. The substrate used was nutrient soil. All plants were cultivated in a greenhouse or artificial climate chamber under controlled temperature and light conditions, maintaining identical light, temperature, humidity, and water and fertilizer management to minimize environmental variation. Three treatment groups were set up: a blank control group (CK), which was regularly watered with the same amount of water; a traditional inducer treatment group (T1), which received a 100 μM methyl jasmonate aqueous solution; and a novel peptide treatment group (T2), which received a 25 nM synthetic SmIDL1 peptide aqueous solution. Each group had at least 10 biological replicates, arranged in a completely randomized block design. Treatments began during the vigorous growth period of *Salvia miltiorrhiza*, administered via foliar spraying every 10 days, for a total of 3-4 treatments. After the final treatment and a period of uniform material accumulation, all plants were harvested. The substrate was carefully shaken off the roots, washed with clean water, and the roots were completely separated. Root samples were first blanched in a 105°C oven for 30 minutes, then dried in a 65°C oven to constant weight. The dry weight of each root was measured using a precision electronic balance. The dried root samples from the same treatment group were mixed, pulverized, and passed through a 60-mesh sieve to prepare a uniform powder sample. Equal amounts of sample were accurately weighed from each treatment group, and the content of the main active ingredient, tanshinone IIA, was determined using a validated high-performance liquid chromatography (HPLC) method. Chromatographic conditions were fixed (e.g., a specific C18 column, a methanol-water mobile phase system, and a detection wavelength of 270 nm). The peak area of ​​the sample was compared with a standard curve plotted using the external standard method to accurately calculate the content (mg / g dry weight). Combining the dry weight data of each root with the measured component content, the yield of tanshinone IIA per plant under each treatment (mg / plant) was calculated.

[0042] Figure 4 clearly shows that different treatments had significant differential effects on the growth and medicinal component accumulation of *Salvia miltiorrhiza*. Specifically, compared with the blank control group (CK, root dry weight 16.0 g / plant, tanshinone IIA content 2.3 mg / g, yield per plant 36.8 mg), the traditional inducer methyl jasmonic acid treatment (T1) increased the tanshinone IIA content to 2.9 mg / g and the yield per plant to 45.0 mg, but its root dry weight (15.5 g / plant) decreased slightly, showing an effect pattern of "improving quality but slightly inhibiting yield". In contrast, the SmIDL1 peptide treatment (T2) of this invention showed a superior synergistic improvement advantage: it not only significantly increased the biomass accumulation of the root system (root dry weight 17.0 g / plant, the highest among the three groups), but also greatly increased the tanshinone IIA content to 3.5 mg / g, thereby driving the yield per plant to a peak of 59.5 mg. This indicates that the SmIDL1 treatment successfully achieved a high-efficiency balance between "increased yield" and "improved quality." The yield of tanshinone IIA per plant increased by approximately 61.7% and 32.2% compared to the CK group and T1 (MeJA) group, respectively. The overall effect was significantly better than that of traditional chemical inducers, highlighting its great potential and application value as a novel biological agent in the targeted improvement of the quality of tanshinone.

[0043] Example 5: Verification of the "Precision Quality Improvement Method in Late Development" under field conditions.

[0044] The study was conducted at the Danshen Shandong base. Three treatments were established: CK (conventional management), T1 (sprayed with 100 µM MeJA, conventional inducer control), and T2 (sprayed with 20 nM SmIDL1, this invention). Both T1 and T2 were applied as a single spray 112 days after transplanting. Yield was measured and components were analyzed.

[0045] The results are as follows:

[0046] Conclusion: The method of this invention achieves comparable or even better yield increase, while the increase in tanshinone IIA content is far greater than that of the MeJA treatment, and there are no adverse reactions in the aboveground parts, with only the lower leaves showing the expected metabolic transformation characteristics. The overall advantages are obvious.

[0047] Example 6: Investigation of Mechanism of Action - Gene Expression Analysis.

[0048] To systematically elucidate the mechanism by which SmIDL1 peptide regulates the specific accumulation of tanshinone at the molecular level, this embodiment designed and implemented a gene expression time-series analysis experiment. Two-year-old potted tanshinone seedlings with consistent growth cycles and stable physiological states were selected as materials and randomly divided into experimental and control groups. The experimental group was foliar sprayed with the optimal working concentration (20 nM) of the SmIDL1 peptide preparation, while the control group was sprayed with an equal volume of blank buffer. A carefully designed sampling strategy was employed: at four key time points after spraying—0 (baseline value before treatment), 3, 12, and 24 hours—mature functional leaves, root tips, and active secondary root tissues from the middle and lower parts of both groups were simultaneously collected. Three independent individuals were taken from each time point as biological replicates. Immediately after sampling, the samples were flash-frozen in liquid nitrogen and stored at -80°C to maximize RNA integrity and ensure the reliability of subsequent analytical data.

[0049] Total RNA was extracted from leaf and root samples using an RNA extraction kit suitable for polysaccharide and polyphenol-rich plant tissues. The integrity, purity, and concentration of the RNA were rigorously checked using agarose gel electrophoresis and a Nanodrop micro spectrophotometer to ensure that the A260 / A280 ratio of all samples was between 1.8 and 2.0. Subsequently, high-quality first-strand cDNA was synthesized using a PrimeScript RT reverse transcription kit with 1 μg of total RNA as a template after strict removal of genomic DNA contamination. All cDNA templates were uniformly diluted and stored at -20°C for later use.

[0050] In quantitative real-time PCR (qRT-PCR), we selected four key functional genes: at the source end (leaf), we selected the sugar transporter gene SmSWEET11 (a key marker representing the ability to export photosynthetic products) and the senescence-related gene SmSAG12 (a classic marker representing the initiation of senescence and nutrient remobilization signals); at the sink end (root), we selected two rate-limiting enzyme genes in the tanshinone synthesis pathway, SmCPS1 (cobamic acid pyrophosphate synthase gene) and SmKSL1 (kaurene synthase gene). qRT-PCR was performed using the SYBR Green Premix Pro Taq HS premixed enzyme system, with three technical replicates for each reaction. The reaction program was: 95℃ pre-denaturation for 30 seconds; followed by 40 cycles of 95℃ for 5 seconds and 60℃ for 30 seconds; finally, melting curve analysis was performed to confirm the uniqueness of the amplified products. Data were analyzed using the comparative Ct value method (2-ΔΔCt method), with the expression level of the control group sample at 0 hours calibrated as 1, and the relative expression levels of each gene at different treatments, tissues, and time points were calculated.

[0051] Figure 5 shows the expression changes of senescence-related genes SmSAG12 and SmSWEET in leaves, as well as key genes for tanshinone synthesis in roots, SmCPS1 and SmKSL1, at different time points after spraying. Based on qRT-PCR time-series analysis data, SmIDL1 treatment produced a systematic and time-series-specific regulatory effect on the expression of key functional genes in *Salvia miltiorrhiza*. Specifically, regarding genes related to leaf senescence and nutrient remobilization, SmSAG12 (a senescence marker gene) began to be upregulated 3 hours after SmIDL1 treatment and continued to rise to 2.4 times that of the control at 24 hours, indicating that senescence-related signaling pathways were effectively activated. Meanwhile, the expression response of the sugar transporter gene SmSWEET was more rapid and intense, reaching 2.4 times that of the control at 3 hours and further increasing to 3.1 times at 24 hours, indicating a significant enhancement in leaf sugar output capacity. In the root tanshinone biosynthesis pathway, the expression of two key enzyme genes, SmCPS and SmKSL1, was also significantly induced: both showed an increase in expression after 3 hours of treatment and continued to increase over time, reaching 2.4-fold and 2.3-fold higher than the control at 24 hours, respectively, and their overall expression trend was significantly higher than their respective control groups. These results indicate that SmIDL1 treatment can rapidly initiate sugar transport and senescence-related signals in leaves at an early stage, thereby synergistically and continuously activating the root tanshinone biosynthesis pathway. This provides a complete transcriptional explanation of the molecular mechanism by which this polypeptide achieves tanshinone-specific accumulation by coordinating the directed enhancement of photosynthetic product output at the "source" end and secondary metabolic flux at the "sink" end.

[0052] Finally, it should be noted that although the above embodiments describe specific implementations of the present invention, they are not intended to limit the invention. Those skilled in the art should understand that these are merely illustrative examples, and the scope of protection of the present invention is defined by the appended claims. All modifications or equivalent substitutions should be included within the scope of protection of the present invention.

Claims

1. A tanshinone SmIDL1 polypeptide, the amino acid sequence of which is shown in SEQ ID NO.

1.

2. The method for preparing the SmIDL1 polypeptide of Danshen as described in claim 1, characterized in that: It is artificially synthesized.

3. The application of the SmIDL1 polypeptide of Danshen according to claim 1 in the preparation of formulations that increase the content of lipid-soluble active ingredients in Danshen.

4. The application as described in claim 3, characterized in that: The active ingredients of tanshinone mentioned above are tanshinone IIA and cryptotanshinone.

5. The application as described in claim 3, characterized in that: The effective working concentration of the tanshinone SmIDL1 polypeptide in the formulation is 1.0 nM ~ 200 nM.

6. The application as described in claim 3, characterized in that: The solvent for the formulation is a weakly acidic buffer solution with a pH of 5.8-6.2, or a buffer system containing the following auxiliary components: energy metabolism synergist: 0.05-0.2 mM adenosine-5'-phosphate; wetting agent: 0.005%-0.01% by volume of plant-derived surfactant.

7. The method for preparing the formulation according to claim 3, characterized in that... Includes the following steps: The peptide SEQ ID NO: 1 was synthesized in a solid phase, purified by HPLC, and then lyophilized. When needed, it was dissolved in the above buffer solution to prepare a working solution, which should be prepared and used immediately.

8. A method for increasing the content of active ingredients in Danshen using the formulation of claim 3, characterized in that: During the early stage of reproductive growth of Salvia miltiorrhiza and before the peak flowering period, the prepared solution is sprayed onto the mature leaves of the lower and middle parts of the plant.

9. The method as described in claim 8, characterized in that... Includes the following steps: It can be applied as a single spray or in two separate sprays. A single spray is applied 110-115 days after transplanting, using an effective working concentration of SmIDL1. A two-spray application is applied first at the budding stage after transplanting, and then 10-15 days after the first application, i.e., before the full bloom stage, using an effective working concentration of SmIDL1.

10. A type of Salvia miltiorrhiza medicinal material obtained by cultivation using the method described in claim 8 or 9.

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