CN-121991248-A - Stability-enhanced plant immunity inducer based on mesoporous silica coupled immune protein and application thereof

Abstract

The invention discloses a stability-enhanced plant immunity inducer based on mesoporous silica coupled immune protein and application thereof, belonging to the technical field of plant disease control. The invention creatively integrates fluorescence visualization and nano-domain stability enhancement technology into a mesoporous silica protein delivery system, and successfully solves the two core problems of difficult tracking and poor stability in research and application of plant immune induced resistance protein. The system can be used as a high-efficiency plant vaccine delivery product, the visual characteristic of the system makes the system a powerful basic research platform, provides a key technical support for optimizing delivery efficiency and clarifying a resistance induction mechanism, and has great application value and market prospect in the green agriculture field.

Inventors

• DING XINHUA
• Lu Chongchong
• DENG MIAOMIAO
• GAO YUAN
• LI ZIMENG
• WANG XIAOWEN

Assignees

• 山东农业大学

Dates

Publication Date

20260508

Application Date

20260126

Claims (9)

1. 1. A stability-enhanced plant immunity inducer based on mesoporous silica coupled immune protein is characterized by being prepared by the following method: Mixing and reacting the surface-modified immune protein or immune protein-fluorescent fusion protein with mesoporous silica nano-particles carrying amino groups or carboxyl groups, so that the immune protein or immune protein-fluorescent fusion protein is stabilized in the pore channels of the mesoporous silica nano-particles in a chemical bonding mode.
2. 2. The plant immune elicitor according to claim 1, wherein the immune protein is selected from chitinase, an exciton protein or an antimicrobial peptide.
3. 3. The plant immunity-inducing antibody according to claim 2, wherein the immune protein is flg22, the amino acid sequence of which is shown in SEQ ID NO.1, and the immune protein-fluorescent fusion protein is flg22-GFP, the amino acid sequence of which is shown in SEQ ID NO. 6.
4. 4. The plant immunity-inducing antibody according to claim 1, wherein the surface modification treatment method of the immune protein-fluorescent fusion protein comprises the steps of uniformly mixing the immune protein-fluorescent fusion protein, NHS and EDC, and reacting for 15-30 minutes in ice in the absence of light.
5. 5. The plant immunity-inducing agent according to claim 1, wherein the mesoporous silica nanoparticle has a particle size of 1 to 1000 nm.
6. 6. Use of the plant immunity-inducing agent according to any one of claims 1 to 5 in at least one of the following (1) - (3): (1) Visual observation of target immune elicitor proteins in plant tissues; (2) Improving the immunity resistance of the plants; (3) Improving the resistance of plants to pathogens.
7. 7. The use according to claim 6, wherein the pathogen is rice bacterial leaf spot.
8. 8. A method for improving disease resistance of a plant, comprising the step of applying the above plant immunity-inducing agent to the plant.
9. 9. The method of claim 8, wherein the plant immune elicitor agent is applied by spraying, watering, or drip irrigation.

Description

Stability-enhanced plant immunity inducer based on mesoporous silica coupled immune protein and application thereof Technical Field The invention relates to the technical field of plant disease control, in particular to a stability-enhanced plant immunity inducer based on mesoporous silica coupled immune protein and application thereof. Background The protein biological pesticide is a biological source pesticide which takes protein or polypeptide as an active ingredient, realizes green prevention and control by activating a plant immune system or directly interfering with the physiological functions of pathogens/pests, and is an important research direction of future pest green prevention and control. At present, the traditional protein pesticide application process has two major bottlenecks, namely firstly, the delivery and release dynamics of the protein in a plant body cannot be observed in real time and intuitively, the optimization and mechanism research of a delivery system are limited, secondly, plant leaves have special structures such as horny layers, waxy layers and epidermal hairs and the like, have barrier effects, seriously obstruct the delivery and permeation of medicaments, the traditional medicament spraying has insufficient physical and chemical stability protection and poor permeability on immune induced protein in the delivery process, a large amount of medicaments are lost in the natural medicament diffusion process, and the sufficient amount of functional proteins cannot be ensured to break through the physical barrier of the plant leaves to reach an action target point, so that the medicament effect is reduced and environmental pollution is caused. The mesoporous silica nanoparticle is used as an excellent drug delivery carrier, has adjustable pore diameter, high specific surface area and easy functionalized surface, and provides an ideal carrier platform for drug embedding and delivery. However, the delivery process of using fluorescent protein to trace immune antigen protein is still limited by a plurality of technical limitations that firstly, the fusion of fluorescent protein and immune antigen protein can change the natural conformation of target protein, thereby affecting the bioactivity, receptor recognition capability and subcellular localization of target protein, so that the tracing result is difficult to fully reflect the real behavior of natural protein, and the fluorescent protein has larger molecular weight, thus the volume and structural complexity of the fusion protein can be obviously increased, and the transport characteristics of small peptide or natural protein can not be accurately simulated. Plant tissues have stronger autofluorescence, and fluorescent proteins can be degraded or fluorescence quenched in plants, so that the signal to noise ratio is reduced, and long-term and stable dynamic observation is difficult to realize. Meanwhile, the fluorescent signal cannot effectively distinguish the "protein still encapsulated by the carrier" from the "released protein", so that the release kinetics are difficult to be accurately resolved. Second, when mesoporous silica is used to deliver immunoelicitor proteins fused to fluorescent proteins, additional challenges are faced with respect to the properties of the carrier. The molecular weight of the fusion protein is obviously increased, and the molecular weight possibly exceeds the pore diameter range of part of mesoporous silica, so that the embedding efficiency is reduced or the fusion protein cannot effectively enter into pore channels, and the loading capacity and the delivery efficiency are affected. The increase in volume of the fusion protein may also alter its diffusion behavior within the mesopores, biasing the release kinetics. In addition, the interaction between the mesoporous silica surface and the fusion protein may be enhanced or reduced, thereby affecting the stability and activity of the protein. In summary, the combination of fluorescent protein markers and mesoporous silica delivery systems still has multiple technical difficulties in the aspects of protein activity maintenance, carrier suitability, loading efficiency, stability, dynamic tracking and the like. Disclosure of Invention Aiming at the prior art, the invention aims to provide a stability-enhanced plant immunity inducer based on mesoporous silica coupled immune protein and application thereof. In order to achieve the above purpose, the invention adopts the following technical scheme: In a first aspect of the invention, a stability-enhanced plant immunity-inducing agent based on mesoporous silica coupled immune protein is provided, and is prepared by the following method: mixing and reacting the surface-modified immune protein or immune protein-fluorescent fusion protein with Mesoporous Silica Nanoparticles (MSNs) carrying amino groups or carboxyl groups, so that the immune protein or immune protein-fluorescent fusion protein is stabilized in