CN-120989127-B - Application of rice OsNAR gene or coded protein thereof in regulation and control of rice blast resistance to rice blast bacteria

CN120989127BCN 120989127 BCN120989127 BCN 120989127BCN-120989127-B

Abstract

The invention discloses application of a rice OsNAR gene or a protein coded by the gene in regulating and controlling rice blast resistance of rice, and relates to the technical field of rice blast control. The CDS nucleotide sequence of the rice OsNAR gene is shown as SEQ ID No. 2. According to the invention, the research shows that the rice gene OsNAR has an important effect in the process of resisting rice blast, the number of lesions on the leaves of the gene deletion mutant is smaller, the gene can be used for screening and obtaining rice strains resisting rice blast, the expression quantity of the basic resistance marker gene in the gene deletion mutant is obviously up-regulated, and the expression of the OsNAR negative regulation basic resistance gene can be used for preventing and treating rice blast.

Inventors

KOU YANJUN
SHI HUANBIN
LIU ZHIQUAN
QIU JIEHUA
CHEN ZHENGTING

Assignees

中国水稻研究所

Dates

Publication Date: 20260508
Application Date: 20250729

Claims (7)

1. The application of the silencing or knocking rice OsNAR1 gene or the protein coded by the gene in improving the rice blast fungus resistance of rice is disclosed, the CDS nucleotide sequence of the rice OsNAR gene is shown as SEQ ID No.2, and the amino acid sequence of the protein coded by the rice OsNAR gene is shown as SEQ ID No. 1.
2. A method for regulating and controlling the resistance of rice to rice blast bacteria is characterized in that OsNAR gene in rice is silenced or knocked out to improve the resistance of the rice to rice blast bacteria when the resistance of the rice to rice blast bacteria is required to be improved, and the CDS nucleotide sequence of the rice OsNAR1 gene is shown as SEQ ID No. 2.
3. The application of the rice OsNAR gene in rice breeding is that rice plants with the rice OsNAR gene silenced or knocked out are screened to obtain rice plant lines resisting rice blast germ, and the CDS nucleotide sequence of the rice OsNAR gene is shown as SEQ ID No. 2.
4. The application of the protein coded by the rice OsNAR gene in rice breeding is that rice plants which are not expressed by the protein coded by the rice OsNAR gene are screened to obtain rice plant lines resistant to rice blast pathogens, and the amino acid sequence of the protein coded by the rice OsNAR gene is shown as SEQ ID No. 1.
5. A construction method of transgenic rice resisting Pyricularia oryzae is characterized in that OsNAR gene in rice is silenced or knocked out, CDS nucleotide sequence of rice OsNAR gene is shown as SEQ ID No. 2.
6. The method for constructing a transgenic rice plant resistant to Pyricularia oryzae according to claim 5, comprising the steps of: (1) Constructing a gene silencing or knockout vector which is a plant expression vector with a sequence for silencing or knockout OsNAR genes; (2) Introducing the gene silencing or knocking-out vector in the step (1) into a rice cell to silence or knock out OsNAR gene, and culturing the transgenic rice plant.
7. The method for constructing a transgenic rice plant resistant to Pyricularia oryzae as claimed in claim 6, wherein the target sequence for gene knockout is shown in SEQ ID No. 3.

Description

Application of rice OsNAR gene or coded protein thereof in regulation and control of rice blast resistance to rice blast bacteria Technical Field The invention relates to the technical field of rice blast control, in particular to application of a rice OsNAR gene or a coded protein thereof in regulating and controlling rice blast resistance. Background Rice is a major food source for more than half of the world population, and its production stability is directly related to global food safety. The rice blast is a serious disease caused by fungi Magnaporthe oryzae, and the rice blast can occur at different positions in different stages of rice growth and development, mainly leaf blast, neck blast and grain blast. The rice blast fungus is mainly propagated in a non-ecological mode in the field, and the infection cycle is completed, so that the global rice production is affected. Novel agents effective in controlling rice blast are currently poorly known. However, by using the disease-resistant gene, the resistance of rice varieties to such pathogens can be significantly improved, thereby reducing yield loss caused by rice blast, for example, the patent applications with publication numbers CN115109797a and CN115109786a all use the disease-resistant gene for improving the resistance of rice to rice blast bacteria. The anti-disease gene is mined and applied to improve the resistance of rice to rice blast, has important practical significance in agricultural production practice, and is also important for coping with challenges possibly appearing in the future. Iron-sulfur cluster (ion-sulfur clusters, fe-S clusters) is an important cofactor commonly existing in organisms and participates in the functions of key enzymes and proteins in various life processes. They exist mainly in two forms of [2Fe-2S ] and [4Fe-4S ] and play an indispensable role in a plurality of biological processes such as electron transfer, gene expression regulation and control, environmental perception and the like. The synthesis of iron-sulfur clusters is a complex and finely tuned process involving multiple steps and specific proteins. Nar1 is an iron/sulfur (Fe/S) containing protein, and cell biology and biochemistry studies identified two magnetically coupled [4Fe-4S ] clusters in Nar1 for yeast and plants. The C-terminal Fe/S cluster of yeast Nar1 is assembled depending on the N-terminal cluster, and the deletion of Nar1 can seriously affect the maturation of all the cytoplasmic and nuclear Fe/S proteins studied, while the mitochondrial Fe/S proteins are not. Similar results have recently been obtained also in human Nar1 homologs IOP1 and Chlamydomonas Nar 1. In Arabidopsis NAR1 is necessary for the function of both gametophytes and synthons. In female gametophytes, NAR1 is necessary for transcriptional activation of certain imprinted genes and is also necessary for proper abiotic stress response in vegetative tissues. However, the biological function of NAR1 in rice is not yet clear. Disclosure of Invention According to the research, the rice OsNAR gene is related to the immune response of rice to rice blast, after the OsNAR gene is overexpressed, the defense capacity of the rice against rice blast pathogens is reduced, the knockout or silencing OsNAR1 gene can improve the disease resistance of the rice against rice blast pathogens, the expression amounts of the basic resistance marker gene OsKS and the OsPR1A in the knockout plants are obviously up-regulated, and the rice blast can be prevented and treated by using the gene as a medicament. The technical scheme of the invention is as follows: The invention provides application of rice OsNAR gene or protein encoded by the gene in regulating resistance of rice to rice blast bacteria, CDS nucleotide sequence of the rice OsNAR gene is shown as SEQ ID No.2, and amino acid sequence of the rice OsNAR gene or protein encoded by the gene is shown as SEQ ID No. 1. The invention also provides application of the rice OsNAR gene or the protein encoded by the gene in preparation of medicines for resisting rice blast bacteria, the CDS nucleotide sequence of the rice OsNAR gene is shown as SEQ ID No.2, and the amino acid sequence of the protein encoded by the rice OsNAR gene is shown as SEQ ID No. 1. Compared with the expression level of the wild type marker, the expression levels of the basic resistance marker gene OsKS and OsPR1A in the knocked-out plants (Osnar-3, osnar 1-17) are obviously up-regulated, which indicates that OsNAR1 negatively regulates the expression of the basic resistance gene. The invention also provides a method for regulating and controlling the resistance of rice to rice blast bacteria, which comprises the steps of over-expressing OsNAR gene in rice when the resistance of rice to rice blast bacteria needs to be reduced, silencing or knocking out OsNAR gene in rice when the resistance of rice to rice blast bacteria needs to be improved, and the CDS nucleotide sequence of OsNA