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CN-121975836-A - Application of Nta04g10180 gene as plant immunity negative regulation factor in improving disease resistance of tobacco

CN121975836ACN 121975836 ACN121975836 ACN 121975836ACN-121975836-A

Abstract

The invention belongs to the technical field of biological agriculture, and particularly relates to application of Nta04g10180 gene and encoding protein thereof in improving black shank resistance of cultivated tobacco. The invention discovers that the Nta04g10180 gene negatively regulates and cultures the resistance of tobacco to phytophthora nicotianae, which is a pathogenic bacterium of tobacco black shank, and experiments prove that silencing the Nta04g10180 gene down regulates the expression of SA (salicylic acid) signal path marker gene NtPR, activates SA-dependent immune response, enhances active oxygen burst and further improves disease resistance, and over-expressing the Nta04g10180 gene makes the tobacco more susceptible to diseases. The invention provides a new target point for breeding of cultivated tobacco disease resistance, and the gene is knocked out or the expression of the gene is reduced in the cultivated tobacco by utilizing CRISPR-Cas9 and other gene editing technologies, so that the disease resistance of the tobacco can be enhanced, and the invention has wide application prospect in the aspect of improving the disease resistance of the tobacco.

Inventors

  • LI QIAN
  • SHI YULONG
  • WANG PENGFEI
  • Cao Jihan
  • MA YU
  • WU JIANG
  • LIU LIU
  • NIU LILI

Assignees

  • 河南省烟草公司南阳市公司

Dates

Publication Date
20260505
Application Date
20251125

Claims (9)

  1. The application of the Nta04g10180 gene in improving the black shank resistance of cultivated tobacco is characterized in that the black shank resistance of the tobacco is improved by silencing the Nta04g10180 gene, and the CDS sequence of the Nta04g10180 gene is shown as SEQ ID NO. 1.
  2. 2. The use according to claim 1, wherein the Nta04g10180 gene is capable of increasing tobacco black shank resistance by modulating the SA signaling pathway.
  3. 3. The use of claim 2, wherein said modulating the SA signaling pathway is up-regulating the expression of SA signaling pathway marker gene NtPR.
  4. 4. The use according to claim 1, wherein said tobacco black shank is induced by phytophthora nicotianae.
  5. 5. The application of the white Nta04g10180 in improving the black shank resistance of cultivated tobacco is characterized in that the expression of the protein Nta04g10180 is reduced or the activity of the protein Nta04g10180 is weakened to improve the black shank resistance of the tobacco, the protein Nta04g10180 is encoded by the Nta04g10180 gene, and the amino acid sequence of the protein Nta04g10180 is shown as SEQ ID NO. 2.
  6. 6. A method for improving black shank resistance of cultivated tobacco is characterized in that in-vivo silencing of Nta04g10180 gene or silencing of specific silencing fragment of Nta04g10180 gene of cultivated tobacco is carried out, CDS sequence of the Nta04g10180 gene is shown as SEQ ID NO. 1, and nucleotide sequence of the specific silencing fragment of the Nta04g10180 gene is shown as SEQ ID NO. 3.
  7. 7. A method of breeding a black shank resistant variety of cultivated tobacco comprising silencing the Nta04g10180 gene or silencing a specific silencing fragment of the Nta04g10180 gene in a plant.
  8. 8. A method for cultivating a cultivated tobacco black shank resistant variety, characterized in that the black shank is caused by phytophthora nicotianae infection.
  9. 9. The method of claim 8, wherein the Nta04g10180 gene silencing plant is obtained by constructing a gene silencing vector, transforming the gene silencing vector into a plant body by an agrobacterium-mediated method.

Description

Application of Nta04g10180 gene as plant immunity negative regulation factor in improving disease resistance of tobacco Technical Field The invention belongs to the technical field of biological agriculture, and particularly relates to application of Nta04g10180 gene and encoding protein thereof in improving black shank resistance of cultivated tobacco. Background Tobacco black shank (Phytophthora infestans) is the primary oomycete disease in the production of cultivated tobacco in China, almost covers all producing areas, and is one of main diseases of cultivated tobacco. The pathogenic bacteria are semi-living nutritive and abnormal mating oomycetes, a large number of sporangia are generated on the surface of infected tissues through sporangium peduncles in the asexual propagation stage, sprout tubes are germinated at a high temperature of 20-25 ℃, and zoospores are released at a low temperature of 10-15 ℃. Zoospores can directly penetrate through leaf tissues or stem tissues, cause lesions after 3-4 days, and spread to adjacent fields through wind power, so that large-area infection is caused. Under proper conditions, phytophthora nicotianae can quickly complete the life cycle, form a large number of zoospores and cause systematic damage to roots, stems and leaves of cultivated tobacco. Phytophthora nicotianae has a high degree of genetic variability, and its genomic dynamic regions include large amounts of effector proteins, which regions may be tolerant of more non-allelic recombination and other forms of structural variation. These features promote increased virulence and other beneficial pathogen changes, which allow phytophthora nicotianae to quickly adapt to new selection pressures generated by modern agriculture, making tobacco black shank difficult to control. Current major tobacco cultivars are generally susceptible to tobacco black shank and lack efficient and available disease resistance gene resources. The main control strategy for phytophthora nicotianae induced diseases at present is the use of bactericides. However, the oomycete disease has a large difference from the physiological and biochemical characteristics of the fungal disease, and the bactericide for preventing and controlling the tobacco black shank is limited, and excessive or improper use is easy to cause the problems of drug resistance, pesticide residue and environmental pollution, so that the related genes for resisting the tobacco black shank are searched and excavated from tobacco genes and utilized, and the method has important significance for realizing the prevention and control of the green tobacco black shank. Disclosure of Invention In order to solve the technical problems, the invention obtains the Nta04g10180 gene of cultivated tobacco through a genetic engineering technology, further researches on the functional genes of the cultivated tobacco, discovers that the Nta04g10180 gene participates in immune response of phytophthora nicotianae which is a pathogenic bacterium of tobacco black shank, constructs an Nta04g10180 gene over-expression plant through genetic means and a genetic engineering technology, discovers that the Nta04g10180 gene over-expression plant weakens resistance to the phytophthora nicotianae, constructs an Nta04g10180 gene silencing plant through an agrobacterium-mediated method, discovers that the Nta04g10180 gene silencing plant strengthens resistance to the phytophthora nicotianae, and provides a new biological resource for improving the black shank resistance of cultivated tobacco. In one aspect, the invention provides application of the Nta04g10180 gene in improving black shank resistance of cultivated tobacco, and silencing of the Nta04g10180 gene improves black shank resistance of the tobacco, wherein a CDS sequence of the Nta04g10180 gene is shown as SEQ ID NO. 1. Further, in the application, the Nta04g10180 gene can improve the tobacco black shank resistance by regulating SA signal pathway. Further, in the application, the modulating SA signaling pathway is up-regulating the expression of SA signaling pathway marker gene NtPR. Further, in the application, the tobacco black shank is caused by phytophthora nicotianae infection. In still another aspect, the invention also provides application of the protein Nta04g10180 in improving black shank resistance of cultivated tobacco, the expression of the protein Nta04g10180 is reduced or the activity of the protein Nta04g10180 is weakened to improve the black shank resistance of the tobacco, the protein Nta04g10180 is encoded by the Nta04g10180 gene, and the amino acid sequence of the protein Nta04g10180 is shown as SEQ ID NO. 2. In still another aspect, the invention provides a method for improving black shank resistance of cultivated tobacco, wherein in the cultivated tobacco, the Nta04g10180 gene is silenced or the specific silencing fragment of the Nta04g10180 gene is silenced, the CDS sequence of the Nta04g10180 gene is shown as SEQ ID NO. 1