Search

CN-121975861-A - Application of reducing expression of negative regulation gene in improving phytophthora sojae resistance

CN121975861ACN 121975861 ACN121975861 ACN 121975861ACN-121975861-A

Abstract

The invention discloses an application of reducing the expression of a negative regulation gene in improving the resistance of phytophthora sojae, belonging to the field of plant genetic engineering. Through researches, the soybean gene GmTIFY a or GmTIFY b has the function of negatively regulating and controlling the soybean phytophthora resistance. Knocking-out GmTIFY a or GmTIFY b, or GmTIFY a and GmTIFY b can enhance the resistance of soybean to phytophthora sojae, and can be applied to the improvement of the disease caused by the phytophthora sojae or the cultivation of soybean varieties resistant to the disease caused by the phytophthora sojae. The soybean cultivar Williams 82 double mutant which does not contain GmTIFY a and GmTIFY b has the advantages that the accumulation amount of phytophthora sojae is reduced by 50-60% compared with that of a wild type, the infection area of a disease spot is reduced by 80-90% compared with that of the wild type, the survival rate is improved by 20-30% compared with that of the wild type, and the resistance of plants to phytophthora sojae can be obviously improved by knocking out GmTIFY a and GmTIFY b. By carrying out genetic engineering improvement on the plant phytophthora root rot resistance genes, important gene resources are provided for plant molecular breeding, and an effective means is provided for high yield of plants.

Inventors

  • ZHAO JINMING
  • Cheng Qican
  • DING WENTAO
  • MEI JIAHAO
  • WANG XIUSHUAI
  • XING HAN
  • GUO NA

Assignees

  • 南京农业大学三亚研究院

Dates

Publication Date
20260505
Application Date
20260409

Claims (9)

  1. 1. An application of reducing the expression of negative regulation genes in improving the resistance of phytophthora sojae, wherein the negative regulation genes are soybean genes GmTIFY a for negative regulation of the resistance of the phytophthora sojae or homologous soybean genes GmTIFY5b or combination of soybean genes GmTIFY a and GmTIFY b, the CDS sequence of the soybean genes GmTIFY a is shown as SEQ ID No.2, and the CDS sequence of the soybean genes GmTIFY b is shown as SEQ ID No. 4.
  2. 2. The use according to claim 1, wherein the genomic sequence of soybean gene GmTIFY a is shown in SEQ ID No.1 and the genomic sequence of soybean gene GmTIFY b is shown in SEQ ID No. 3.
  3. 3. The use of claim 1 or 2, wherein the reduction of expression of the negative regulatory gene is a knock-out of soybean gene GmTIFY a, or GmTIFY b, or a combination of GmTIFY a and GmTIFY b using a CRISPR/CAS 9 recombinant knock-out vector.
  4. 4. An application of reducing the expression of negative regulation gene in improving the disease caused by soybean phytophthora sojae resistance, wherein the negative regulation gene is soybean gene GmTIFY a for negative regulation of soybean phytophthora sojae resistance, or soybean gene GmTIFY b homologous to the negative regulation gene, or soybean gene GmTIFY a and GmTIFY b are combined, the CDS sequence of soybean gene GmTIFY a is shown as SEQ ID No.2, and the CDS sequence of soybean gene GmTIFY b is shown as SEQ ID No. 4.
  5. 5. The use according to claim 4, wherein the genomic sequence of soybean gene GmTIFY a is shown in SEQ ID No.1 and the genomic sequence of soybean gene GmTIFY b is shown in SEQ ID No. 3.
  6. 6. The use according to claim 4 or 5, wherein the reduction of expression of negative regulatory genes is in particular the knockdown of soybean gene GmTIFY a, or GmTIFY b, or a combination of GmTIFY a and GmTIFY b using CRISPR/CAS9 recombinant knock-out vectors.
  7. 7. Use of a negative regulatory gene which is a soybean gene GmTIFY a which negatively regulates resistance to phytophthora sojae, or a soybean gene GmTIFY b which is homologous to the negative regulatory gene, or a combination of soybean genes GmTIFY a and GmTIFY b, wherein the CDS sequence of soybean gene GmTIFY a is shown as SEQ ID No.2, and the CDS sequence of soybean gene GmTIFY b is shown as SEQ ID No.4, for reducing expression of the negative regulatory gene in breeding soybean varieties which are resistant to diseases caused by phytophthora sojae.
  8. 8. The use according to claim 7, wherein the genomic sequence of soybean gene GmTIFY a is shown in SEQ ID No.1 and the genomic sequence of soybean gene GmTIFY b is shown in SEQ ID No. 3.
  9. 9. The use according to claim 7 or 8, wherein the reduction of expression of negative regulatory genes is in particular the knockdown of soybean gene GmTIFY a, or GmTIFY b, or GmTIFY a in combination with GmTIFY b, using CRISPR/CAS9 recombinant knock-out vector.

Description

Application of reducing expression of negative regulation gene in improving phytophthora sojae resistance Technical Field The invention belongs to the field of plant genetic engineering, and particularly relates to an application of reducing expression of a negative regulation gene in improving phytophthora sojae resistance. Background Soybean (Glycine max (l.) merr) is an important grain and oil cash crop in our country, and is one of the main sources for obtaining high-quality vegetable proteins by humans. Soybean phytophthora capsici (Phytophthora stem and root rot, PRR) is one of the core factors threatening the soybean productivity in our country, and the disease was listed in the name of one kind of crop diseases and insect pests by the agricultural rural area in 2023. Soybean phytophthora disease generally causes 15% -30% yield reduction, the yield loss of high-sense varieties can reach 50% -70%, and the retransmission area is even in a dead state. The soybean phytophthora in China has the annual average occurrence area exceeding tens of mu, the soybean phytophthora (Phytophthora sojae) is widely distributed in China, the secreted pathogenic effector is various, and the effector sequence is rapidly mutated under the selection pressure of a host, so that the resistance of the main cultivated variety is easily lost. In face of the obvious defect of the current prevention and control measures, the key genes for resisting soybean phytophthora root rot are mined, disease-resistant materials are created, and then new disease-resistant varieties are cultivated, so that the method is the most fundamental and economical way for controlling the diseases. The gene editing technology cuts target sites of target genes to form DNA double-strand breaks, and realizes mutation of the target genes by using a DNA repair mechanism of a receptor. The CRISPR/Cas9 editing technology has the advantages of high editing efficiency, low cost and the like, and becomes the most widely applied gene editing system at present. Therefore, on the basis of conventional control, the disease resistance of soybeans is improved by utilizing molecular breeding means such as gene editing and the like, and the method is one of the most economical and effective control measures. Disclosure of Invention The invention aims to provide an application of reducing the expression of a negative regulation gene in improving the resistance of phytophthora sojae, and provides a soybean gene or a gene combination for negative regulation of the resistance of phytophthora sojae, wherein the negative regulation gene or the gene combination is knocked out in plants, so that the resistance of soybean to the phytophthora sojae can be improved, and the soybean variety is improved. The aim of the invention is mainly realized by the following technical scheme: The invention discloses a gene for negative regulation of phytophthora sojae resistance, which comprises a soybean gene GmTIFY a, or a soybean gene GmTIFY b homologous to the soybean gene GmTIFY a, or a combination of the soybean gene GmTIFY a and GmTIFY b, wherein the CDS sequence of the soybean gene GmTIFY a is shown as SEQ ID No.2, the genome sequence of GmTIFY5a is shown as SEQ ID No.1, the CDS sequence of the soybean gene GmTIFY b is shown as SEQ ID No.4, and the genome sequence of GmTIFY b is shown as SEQ ID No. 3. The invention discloses an application of soybean gene GmTIFY a for reducing the negative regulation genes including negative regulation of phytophthora sojae resistance or soybean gene GmTIFY b homologous to soybean gene GmTIFY a or soybean gene GmTIFY a and GmTIFY b in improving the phytophthora sojae resistance, in particular to an application of a CRISPR/CAS 9 recombinant knockout vector to knockout soybean gene GmTIFY5a or GmTIFY5b or GmTIFY a and GmTIFY b. The invention also discloses an application of the soybean gene GmTIFY a for reducing the negative regulation genes including the negative regulation of the phytophthora sojae resistance or the soybean gene GmTIFY b homologous to the soybean gene GmTIFY a or the expression of the combination of the soybean genes GmTIFY a and GmTIFY b in improving the disease caused by the phytophthora sojae resistance of soybean. Specifically, the CRISPR/CAS9 recombinant knockout vector is adopted to knock out soybean gene GmTIFY a, gmTIFY b, or a combination of GmTIFY a and GmTIFY b. The invention also discloses an application of the soybean gene GmTIFY a for reducing the negative regulation genes including the resistance of the soybean phytophthora sojae or the soybean gene GmTIFY b homologous to the soybean gene GmTIFY a or the soybean gene GmTIFY a and GmTIFY b in the cultivation of soybean varieties resistant to diseases caused by the soybean phytophthora sojae, in particular to the knockout of the soybean gene GmTIFY a or GmTIFY5b or the combination of GmTIFY a and GmTIFY5b by adopting a CRISPR/CAS 9 recombinant knockout vector. The invention