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CN-122011211-A - Plant immune elicitor based on rice blast fungus MoRCP protein and application thereof

CN122011211ACN 122011211 ACN122011211 ACN 122011211ACN-122011211-A

Abstract

The invention discloses a plant immune elicitor based on rice blast fungus MoRCP protein and application thereof, and the scheme discovers and proves that the rice blast fungus CFEM protein MoRCP1 can be used as an elicitor to induce rice blast resistance of rice, and provides a brand new view and tool for plant immune research. Meanwhile, the scheme also preliminarily reveals the molecular mechanism of the protein excited immunity by rice receptor kinase, scientific basis is firm, the scheme adopts escherichia coli recombinant expression protein for foliage spraying, the method is simple and convenient, the disease severity after rice blast fungus inoculation can be obviously reduced, the prevention effect is clear, the scheme also takes green and environment-friendly into consideration, the scheme strategy utilizes a plant autoimmune system, the problems of chemical pesticide residue and drug resistance are avoided, and the sustainable development concept is met.

Inventors

  • LI YA
  • CHEN SONGBIAO
  • WANG ZONGHUA
  • LU GUODONG
  • CHEN JING
  • WU QIANFEI
  • CHEN JIA
  • WU YUTING

Assignees

  • 福建农林大学

Dates

Publication Date
20260512
Application Date
20260213

Claims (10)

  1. 1. A recombinant fusion protein having rice blast-inducing activity, comprising a rice blast-inducing protein MoRCP of the family of pathogenic rice bacteria (Magnaporthe oryzae) CFEM, wherein the amino acid sequence of protein MoRCP1 is the sequence set forth in SEQ ID No.1, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 98% or 99% sequence identity to the sequence set forth in SEQ ID No. 1.
  2. 2. The recombinant fusion protein of claim 1, further comprising a heterologous tag protein.
  3. 3. The recombinant fusion protein of claim 2, wherein the heterologous tag protein is maltose binding protein MBP.
  4. 4. A polynucleotide for encoding a recombinant fusion protein according to any one of claims 1 to 3, characterized in that it comprises a nucleotide sequence for encoding said protein MoRCP, said nucleotide sequence being shown as SEQ ID No. 2.
  5. 5. A method for preparing a recombinant fusion protein according to any one of claims 1 to 3, comprising the steps of: (1) Cloning the polynucleotide for encoding the fusion protein to an expression vector to construct a recombinant expression plasmid; (2) Transforming the recombinant expression plasmid into competent cells of escherichia coli to obtain recombinant engineering bacteria; (3) Inducing the recombinant engineering bacteria to express the recombinant fusion protein; (4) And (3) splitting the thalli, and purifying by affinity chromatography to obtain the recombinant fusion protein.
  6. 6. The method according to claim 5, wherein in the step (4), the affinity chromatography is affinity chromatography using an amylose resin.
  7. 7. A method for inducing rice to develop resistance to rice blast, comprising applying the recombinant fusion protein of claim 1 to rice plants.
  8. 8. The method of claim 7, wherein the application mode is foliar spray; the recombinant fusion protein is administered at a concentration of 2 μm to 10 μm; the timing of the application is 24 hours to 72 hours before the rice is inoculated or infected with rice blast bacteria.
  9. 9. Use of a recombinant fusion protein according to any one of claims 1 to 3 for the preparation of a formulation for controlling rice blast.
  10. 10. A formulation for controlling rice blast, characterized in that it comprises the recombinant fusion protein according to any one of claims 1 to 3 as an active ingredient, and an agriculturally acceptable carrier.

Description

Plant immune elicitor based on rice blast fungus MoRCP protein and application thereof Technical Field The invention relates to the field of agricultural biotechnology and plant protection, in particular to a plant immunity stimulator based on rice blast fungus (Magnaporthe oryzae) MoRCP protein and application thereof, wherein the application is application of the plant immunity stimulator in inducing innate immunity of plants (especially rice) and preventing and controlling diseases, and more particularly, the invention relates to a novel function of rice blast fungus CFEM family protein MoRCP1 as a plant mode triggering immunity (PTI) activator and application of the novel function in green prevention and control of rice blast. Background Rice blast is a destructive fungal disease caused by rice blast bacteria, which seriously threatens global rice production and grain safety. At present, the prevention and control of the diseases still highly depend on chemical bactericides, but the long-term use of the chemical bactericides is easy to cause the pathogenic bacteria to generate drug resistance, and potential risks are formed for ecological environment and food safety. Therefore, developing new, environmentally friendly strategies for green control based on the plant's autoimmune system has become an important direction for sustainable development of agriculture. Plant immune elicitors are a class of substances that can mimic pathogen-associated molecular patterns (PAMPs) or damage-associated molecular patterns (DAMP) and systematically boost plant disease resistance by activating pattern-triggered immunity (PTI) in plants. The utilization of the elicitor to induce plant resistance, which is equivalent to "vaccinating" plants, is one of the effective ways to achieve green control of diseases. At present, several microbial elicitors have been reported and used, such as Harpin proteins derived from bacteria, fungal chitooligosaccharides, and the like. However, the existing excitons still have the limitations of insufficient effect stability, relatively narrow action spectrum, higher production cost and the like. Therefore, the novel, efficient and well-defined action mechanism plant immune elicitor is developed, and has important significance for enriching the green prevention and control tool box. The cysteine-rich CFEM (Common IN SEVERAL Fungal Extracellular Membrane proteins) domain is a unique class of extracellular protein structural motifs that are commonly associated with fungal pathogenicity, signal transduction, and host interactions. In Pyricularia oryzae, the genome encodes a total of 21 CFEM domain-containing proteins, but until now, their functional studies have been very limited. The prior public reports have focused on only a few members. For example, CFEM protein Pth11 has been demonstrated to act as a G protein-coupled receptor, directly involved in pathogenic processes of pathogenic bacteria by regulating the formation of bacterial attachment cells and development of invasive structures. The function of most other CFEM proteins, especially the role played in plant-pathogen interactions, is not known. Notably, current research on the rice blast CFEM protein is focused on its function as a causative agent, mainly from the viewpoint of pathogenic bacteria. In the prior art, there is no publication or patent showing that any CFEM protein of Pyricularia oryzae is recognized by plants and acts as a PAMP agonist to activate the plant's innate immune response, thereby developing resistance to the pathogenic bacteria themselves. This cognitive gap has led CFEM to proteins that have long been considered merely "targets" for disease management, rather than available "resources". The scheme of the invention is completed, and the inherent cognition is broken through for the first time. The inventors found from comparative studies that the CFEM protein-deleted strain did not have a significant reduction in pathogenicity, unlike typical pathogenic properties. More unexpectedly, the inventor finds that the protein can be identified by rice receptor kinase (RLK), effectively activates strong PTI reaction, and induces rice to generate obvious resistance to subsequent rice blast bacteria infection. Based on its first discovered CFEM family protein with plant-induced resistance, the inventors named "MoRCP1" (MagnaportheoryzaeResistance-InducingCFEMProtein). The discovery reveals a brand new fungus-source PAMP, and provides brand new material basis and theoretical basis for developing plant immune activators based on pathogen derivatives. Disclosure of Invention In view of the above, the invention aims to provide a plant immune elicitor based on rice blast fungus MoRCP protein and application thereof, and the scheme is a brand-new fungus-source plant immune elicitor and application method thereof, which aims to solve the problems of limited elicitor sources, insufficient action mechanism and the like in the ex