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CN-120944912-B - NtGCN1 gene and application thereof in regulating and controlling chloride ion content of tobacco leaves

CN120944912BCN 120944912 BCN120944912 BCN 120944912BCN-120944912-B

Abstract

The invention discloses NtGCN gene and application thereof in regulating and controlling chloride ion content of tobacco leaves, and relates to the technical field of biology, wherein an amino acid sequence coded by the NtGCN gene is shown as SEQ ID NO. 2. According to the invention, through constructing a plant transformation vector and introducing the plant transformation vector into the Chinese tobacco 100, a NtGCN gene mutant material and an over-expression material are obtained, and analysis shows that the increase of NtGCN1 gene expression quantity can obviously improve the content of chloride ions in tobacco, and NtGCN gene expression quantity is reduced, so that the content of chloride ions in tobacco is also obviously reduced. The gene is used as a target, so that the tobacco germplasm with low chlorine content is cultivated, the tobacco quality is improved, and the gene has important significance.

Inventors

  • HAN HUIGE
  • WU ZHAOHUI
  • HU MENGMENG
  • XU WENZHENG
  • LIU QIAOZHEN
  • SONG YANDAN
  • WANG MENGMENG
  • ZHANG RUI

Assignees

  • 河南省农业科学院烟草研究所

Dates

Publication Date
20260508
Application Date
20250917

Claims (8)

  1. The application of the NtGCN1 gene or NtGCN protein in regulating and controlling the chloride ion content of tobacco is characterized in that the amino acid sequence coded by the NtGCN gene is shown as SEQ ID NO.2, and the amino acid sequence of the NtGCN protein is shown as SEQ ID NO. 2; the regulation is positive regulation.
  2. 2. The use of NtGCN gene or NtGCN protein according to claim 1 for regulating and controlling tobacco chloride ion content, wherein the nucleotide sequence of NtGCN gene is shown as SEQ ID No. 1.
  3. 3. The application of the biological material for reducing NtGCN gene expression quantity in regulating and controlling tobacco chloride ion content is characterized in that the biological material is an expression cassette, a vector or a host cell, the regulation is negative regulation, and the amino acid sequence coded by the NtGCN gene is shown as SEQ ID NO. 2.
  4. 4. The use of the biomaterial for reducing NtGCN gene expression level according to claim 3 for regulating and controlling tobacco chloride ion content, wherein the nucleotide sequence of NtGCN1 gene is shown as SEQ ID No. 1.
  5. 5. The application of the biological material for reducing NtGCN gene expression quantity in tobacco breeding is characterized in that the breeding direction is to cultivate germplasm with low chloride ion content, the biological material is an expression box, a vector or a host cell, and the amino acid sequence coded by the NtGCN gene is shown as SEQ ID NO. 2.
  6. 6. The use of the biological material for reducing NtGCN gene expression in tobacco breeding according to claim 5, wherein the nucleotide sequence of NtGCN gene is shown as SEQ ID No. 1.
  7. 7. A method for improving tobacco quality is characterized in that a biological material for reducing NtGCN gene expression level is utilized to construct tobacco germplasm with low chloride ion content; The biological material is an expression cassette, a vector or a host cell, and the amino acid sequence coded by the NtGCN gene is shown as SEQ ID NO. 2.
  8. 8. The method for improving tobacco quality according to claim 7, wherein the nucleotide sequence of NtGCN gene is shown as SEQ ID NO. 1.

Description

NtGCN1 gene and application thereof in regulating and controlling chloride ion content of tobacco leaves Technical Field The invention relates to the technical field of biology, in particular to NtGCN gene and application thereof in regulating and controlling chloride ion content of tobacco leaves. Background Chloride ion is an important micronutrient and accumulates in large quantities in many plants. Tobacco is a chlorine-rich plant with leaf chloride ion concentration up to 50 mg.g -1, but its underlying physiological mechanism is not yet known. The chlorine content in the tobacco plays an important role in the growth of the tobacco and the quality of products thereof, and the excessive chlorine content can cause the thickening, the crispness and the curling of leaf edges, the dark green leaf color (the green loss of lower leaf), the poor smell generated during storage due to the enhanced hygroscopicity, the increased accumulation of starch in the tobacco, the reduced content of reducing sugar and the strange taste, and the chloride ions in the tobacco can also obviously reduce the combustibility and cause the black ash flameout phenomenon. At present, the measures adopted for reducing the chlorine content in tobacco leaves mainly comprise selecting soil with low chlorine background, applying potassium sulfate fertilizer to replace potassium chloride and the like, but the effect is poor and limited by soil regions. Therefore, how to reduce the chlorine content in tobacco is a problem that needs to be solved by those skilled in the art. Disclosure of Invention In view of this, the invention provides NtGCN gene and its application in regulating and controlling chloride ion content in tobacco leaf. In order to achieve the above purpose, the present invention adopts the following technical scheme: NtGCN1 gene, wherein the amino acid sequence coded by NtGCN gene is shown as SEQ ID NO. 2. As a preferable technical scheme, the nucleotide sequence of the NtGCN gene is shown as SEQ ID NO. 1. It is a further object of the present invention to provide NtGCN a protein, the amino acid sequence of said NtGCN a protein is shown as SEQ ID NO. 2. It is still another object of the present invention to provide the use of the NtGCN gene described above, or the NtGCN protein described above, for regulating the chloride ion content of tobacco. It is yet another object of the present invention to provide a biomaterial that reduces NtGCN gene expression levels, which is an expression cassette, vector, or host cell. Wherein the vector comprises an expression vector and a cloning vector. The host cell includes a microbial cell or a plant cell (preferably a non-reproductive plant cell). It is a further object of the present invention to provide the use of the above biological material for regulating the chloride ion content of tobacco. It is a further object of the present invention to provide the use of the above biological material in tobacco breeding in the direction of breeding low chloride ion content germplasm. It is a further object of the present invention to provide a method for improving tobacco quality, utilizing the above-described biological materials to construct tobacco germplasm with low chloride ion content. The invention has the beneficial effects that NtGCN genes and NtGCN1 proteins are obtained through screening and the application of the NtGCN genes and NtGCN proteins in regulating and controlling the chloride ion content of tobacco leaves. The invention constructs a plant transformation vector and introduces the plant transformation vector into the tobacco 100 to obtain a NtGCN gene mutant material and an over-expression material, and analysis shows that the increase of NtGCN1 gene expression level can obviously improve the content of chloride ions in tobacco, and the NtGCN gene expression level is reduced, so that the content of chloride ions in tobacco is also obviously reduced. The gene is used as a target, so that the tobacco germplasm with low chlorine content is cultivated, the tobacco quality is improved, and the gene has important significance. Drawings In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the embodiments or the description of the prior art will be briefly described below, and it is obvious that the drawings in the following description are only embodiments of the present invention, and that other drawings can be obtained according to the provided drawings without inventive effort for a person skilled in the art. FIG. 1 shows the phenotype of tobacco cultivated by high concentration of chloridion, and the wilting of tobacco plants and the decay of root systems appear. FIG. 2 shows the phenotype of 0-45mM chloride cultured tobacco, wherein T0-T5 corresponds to 0, 5, 15, 25, 35 and 45mM concentrations, respectively. FIG. 3 shows the phenotype of 0-25mM chloride cultured tobac