CN-121992009-A - Application of rape NLR type gene cluster in enhancing rice resistance

Abstract

The invention discloses application of rape NLR type gene cluster in enhancing rice resistance. The rape NLR protein pair consists of the proteins 1) and 2), wherein 1) BnNLR1.1 has an amino acid sequence shown as a sequence 1 in a sequence table, and 2) BnNLR1.2 has an amino acid sequence shown as a sequence 3 in the sequence table. By adopting the rape NLR protein pair disclosed by the invention to construct a double-gene heterologous transformation system, the innate immune response of rice can be effectively stimulated, so that the disease resistance of the rice is enhanced. The invention has good genetic stability and disease-resistant effect, provides a new technical path for cultivating new rice varieties with high yield, high quality and disease resistance, and has important agricultural application value and popularization prospect.

Inventors

• WEI JIA
• MA WENDI
• TONG XIN

Assignees

• 中国农业大学

Dates

Publication Date

20260508

Application Date

20251203

Claims (5)

1. 1. An application of a rape NLR protein pair and a coding gene thereof in enhancing disease resistance of rice, wherein the rape NLR protein pair consists of the proteins in the following 1) and 2): 1) BnNLR1.1 has an amino acid sequence shown as a sequence 1 in a sequence table; 2) The BnNLR1.2 has an amino acid sequence shown as a sequence 3 in a sequence table.
2. 2. The use according to claim 1, wherein, The nucleotide sequence of the coding gene of BnNLR1.1 is shown as a sequence 2 in a sequence table; The nucleotide sequence of the coding gene of BnNLR1.2 is shown as a sequence 4 in a sequence table.
3. 3. The use according to claim 1, wherein the disease resistance is rice blast resistance.
4. 4. A method for enhancing resistance of rice by utilizing rape NLR type gene cluster comprises the steps of transferring the encoding genes of BnNLR1.1 and BnNLR1.2 into rice to obtain transgenic rice co-expressed by the BnNLR1.1 and BnNLR1.2, namely the transgenic rice with improved disease resistance, wherein the nucleotide sequence of the encoding gene of the BnNLR1.1 is shown as a sequence 2 in a sequence table, and the nucleotide sequence of the encoding gene of the BnNLR1.2 is shown as a sequence 4 in the sequence table.
5. 5. The use according to claim 4, wherein the disease resistance is rice blast resistance.

Description

Application of rape NLR type gene cluster in enhancing rice resistance Technical Field The invention belongs to the technical field of genetic engineering, and particularly relates to application of a rape NLR type gene cluster in enhancing rice resistance. Background The blast caused by the infection of rice with Pyricularia oryzae is the most destructive rice disease, the first of ten fungal diseases (Dean et al 2012). Pathogenic bacteria infection seriously affects normal plant physiological metabolism of rice, obviously inhibits the nutrition growth and reproductive development process of the rice, and finally leads to sharp reduction of seed yield and quality degradation. This persistent agricultural bio-threat not only affects regional food supplies, but also creates a serious challenge to the global food safety system. Under the background, the breeding of high-quality rice germplasm resources with durable resistance has become a strategic measure in the field of plant pathology prevention and control. The core strategy of modern crop genetic improvement focuses on deep excavation and innovative utilization of resistance gene resources, particularly attaches importance to identification of resistance genes, and the genetic locus with multiple disease resistance functions not only can enhance autoimmune response of crops, but also can provide a molecular breeding foundation for constructing a sustainable disease comprehensive treatment system. Through integrating genomics and molecular design breeding technology, the resistance gene resource is converted into actual productivity, so that the stable operation of a grain system is ensured. The plant and pathogenic microorganism form a multi-level innate immune defense system through long-term co-evolution to inhibit infection of pathogenic bacteria. The defense system is largely divided into two basic immune modes, primary immune response (PTI) based on pathogen-associated molecular patterns (pathogen-associated molecular patterns, PAMPs) and secondary immune response (Effector-TRIGGERED IMMUNITY, ETI) where effectors trigger immunity, depending on the differences in molecular recognition mechanisms. PTI is used as a basic defense layer, and a pattern recognition receptor (pattern recognition receptors, PRRs) positioned by a cell membrane recognizes immune processes of a pathogen-related molecular pattern (pathogen-associated molecular patterns, PAMPs) or a damage-related molecular pattern (damage-associated molecular patterns, DAMPs), so that early defense reactions such as calcium ion inflow and active oxygen burst are activated. However, in the continuous co-evolution of disease systems, although plant PTI successfully resists attack by most pathogens, a few adaptive pathogens (adapted pathogen) utilize evolved effectors (effectors) to suppress plant PTI responses, again effecting infestation of plants. ETI is a highly effective defense mechanism of plants against both biotrophic and semi-biotrophic pathogens. When the effector secreted by the pathogen within the plant cell is recognized by Nucleotide binding leucine-rich repeat receptors (NLRs), the ETI signaling pathway is activated, triggering an allergic cell death response (HYPERSENSITIVE RESPONSE, HR), thereby effectively inhibiting the progress of pathogen infection. Classical NLRs proteins have a ternary domain configuration, including the N-terminal coiled coil (Coiled-coil, CC), the RPW 8-like coiled coil (CC R) or the Toll/interleukin-1 receptor (TIR) domain, the middle APAF-1 protein, the nucleotide binding domain (NB-ARC) common to specific plant disease resistance gene products and nematode CED-4 proteins, and the C-terminal leucine-rich repeat (Leucine-RICH REPEAT, LRR) domain (Jones and Dangl 2006). The part NLRs proteins function in complementary pairs, achieving synergy through negative regulatory mechanisms (e.g., the rice RGA4/RGA5 system; xi et al 2022) or positive regulatory mechanisms (e.g., the Brassica napus BnRPR/BnRPR system; MERMIGKA ET al 2023). The negative regulatory pair is called an helper/sensor type NLR protein pair, wherein the sensor type NLR carries an integration domain (INTEGRATED DOMAINS, IDs), a molecular marker capable of specifically recognizing pathogen effectors, and thus activates helper-type NLR-dependent effector-triggered immunity (ETI). The positive regulatory mechanism involves the co-initiation of an immune response by the two NLR members through synergy. Disclosure of Invention The invention identifies a novel NLR protein pair BnNLR1.1/BnNLR1.2 in Brassica napus cv. Westar. Based on the above, the invention provides an application of a rape NLR protein pair and a coding gene thereof in enhancing rice disease resistance, wherein the rape NLR protein pair consists of the proteins as described in the following 1) and 2): 1) BnNLR1.1 has an amino acid sequence shown as a sequence 1 in a sequence table; 2) The BnNLR1.2 has an amino acid sequence shown