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CN-122012607-A - Gene PbTCP for regulating and controlling pear fruit stone cells

CN122012607ACN 122012607 ACN122012607 ACN 122012607ACN-122012607-A

Abstract

The invention discloses a gene PbTCP for regulating and controlling pear fruit stone cells, wherein the CDS sequence of the gene is shown as SEQ ID NO.1, and the coded protein sequence is shown as SEQ ID NO. 2. The gene construction over-expression vector is injected into the Dangshan pear fruits, compared with the injection of empty control parts, the content of stone cells and lignin is obviously reduced, the gene construction over-expression vector is introduced into the arabidopsis, the secondary cell walls of the duct cells of the PbTCP transgenic arabidopsis obtained are obviously thinned, and the accumulation of lignin in stems is obviously reduced. The discovery of the gene provides a new gene resource for fruit quality breeding, and is an important candidate gene for future genetic engineering improvement fruit quality breeding.

Inventors

  • TAO SHUTIAN
  • HE JIANAN
  • ZHANG SHAOLING
  • QI KAIJIE
  • XIE ZHIHUA

Assignees

  • 南京农业大学三亚研究院
  • 南京农业大学

Dates

Publication Date
20260512
Application Date
20260416

Claims (9)

  1. 1. Use of gene PbTCP13 in (A1) - (A3) as follows: (A1) Application of reducing lignin content in pear fruit or arabidopsis thaliana; (A2) Use in the preparation of a product for reducing lignin content in pear fruits or arabidopsis thaliana; (A3) Application in breeding for reducing lignin content in pear fruits or arabidopsis; The CDS sequence of the gene PbTCP is shown as SEQ ID NO. 1; The application is realized by transferring the gene PbTCP to pear fruit or Arabidopsis thaliana for overexpression.
  2. 2. Use of a protein encoded by the gene PbTCP according to claim 1 in (A1) to (A3) as follows: (A1) Application of reducing lignin content in pear fruit or arabidopsis thaliana; (A2) Use in the preparation of a product for reducing lignin content in pear fruits or arabidopsis thaliana; (A3) Application in breeding for reducing lignin content in pear fruits or arabidopsis; The amino acid sequence of the protein is shown as SEQ ID NO. 2; The application is realized by transferring the gene PbTCP to pear fruit or Arabidopsis thaliana for overexpression.
  3. 3. Use of a recombinant expression vector and/or a transient expression vector comprising the gene PbTCP according to claim 1 in (A1) - (A3) as follows: (A1) Application of reducing lignin content in pear fruit or arabidopsis thaliana; (A2) Use in the preparation of a product for reducing lignin content in pear fruits or arabidopsis thaliana; (A3) Application in breeding for reducing lignin content in pear fruits or arabidopsis; The CDS sequence of the gene PbTCP is shown as SEQ ID NO. 1; The application is realized by transferring the gene PbTCP to pear fruit or Arabidopsis thaliana for overexpression.
  4. 4. The use according to claim 3, wherein the backbone vector of the recombinant expression vector is pCAMBIA1300-GFP.
  5. 5. Use of a recombinant bacterium comprising the gene PbTCP according to claim 1 in (A1) to (A3) as follows: (A1) Application of reducing lignin content in pear fruit or arabidopsis thaliana; (A2) Use in the preparation of a product for reducing lignin content in pear fruits or arabidopsis thaliana; (A3) Application in breeding for reducing lignin content in pear fruits or arabidopsis; The CDS sequence of the gene PbTCP is shown as SEQ ID NO. 1; The application is realized by transferring the gene PbTCP to pear fruit or Arabidopsis thaliana for overexpression.
  6. 6. A method for reducing lignin content of pear fruits, which is characterized in that the method is realized by transferring a gene PbTCP to pear fruits for over-expression, and the CDS sequence of the gene PbTCP is shown as SEQ ID NO. 1.
  7. 7. The method of claim 6, wherein the method is performed by increasing the expression level of gene PbTCP in pear.
  8. 8. A method for reducing lignin content in arabidopsis, which is characterized in that the method is realized by transferring a gene PbTCP to arabidopsis for overexpression, and the CDS sequence of the gene PbTCP is shown as SEQ ID NO. 1.
  9. 9. The method according to claim 8, wherein the method is carried out by increasing the expression level of gene PbTCP in Arabidopsis thaliana.

Description

Gene PbTCP for regulating and controlling pear fruit stone cells Technical Field The invention relates to the fields of plant molecular biology and fruit tree genetic breeding, in particular to a key gene PbTCP for regulating and controlling stone cell formation of pear fruits and application of the gene in pear fruit quality improvement. Background Pear is one of important fruit tree species in Jiangsu province, pear often forms stone cells due to the lignification of cells, and the special lignification cells which significantly affect the quality in the pear form development process, wherein the development process comprises the steps of gradually thickening the secondary cell walls of parenchyma cells to form hard cell walls, and lignin is the main component of the secondary walls, so that the metabolism of lignin and the formation of stone cells have a dense and inseparable relationship (Tao et al 2009). However, the regulatory network behind it has not been fully resolved due to the complexity of the stone cell formation mechanism. Therefore, the key factors for further excavating the stone cell formation are of great significance for accelerating the improvement of pear quality. Disclosure of Invention Aiming at the defects of the prior art, the invention aims to provide a key gene for regulating and controlling the formation of pear fruit stone cells. The gene is obtained by separating and cloning a variety 'Dangshan pear' (Pyrus bretschneideri) with high stone cell content, the applicant names PbTCP, the CDS sequence of the gene is shown as SEQ ID NO.1, and the corresponding protein sequence of the gene is shown as SEQ ID NO.2 of a sequence table. The discovery of the gene can provide new insight for the quality improvement of pear fruit. It is a further object of the present invention to provide an application of the aforementioned gene PbTCP and its related biological material. The gene is constructed into an over-expression vector, and is introduced into Arabidopsis through agrobacterium-mediated genetic transformation, and the obtained transgenic material is verified by biological functions, so that the PbTCP gene cloned by the invention has the functions of reducing lignin accumulation and thickening secondary cell walls. The aim of the invention is achieved by the following technical scheme: in a first aspect, the present invention provides the use of gene PbTCP13 in (A1) - (A3) as follows: (A1) Application of reducing lignin content in plants; (A2) Use in the preparation of a product for reducing lignin content in plants; (A3) Use in breeding to reduce lignin content in plants; The CDS sequence of the gene PbTCP is shown as SEQ ID NO. 1. In a second aspect, the present invention also provides the use of a protein encoded by gene PbTCP in (A1) - (A3) as follows: (A1) Application of reducing lignin content in plants; (A2) Use in the preparation of a product for reducing lignin content in plants; (A3) Use in breeding to reduce lignin content in plants; The amino acid sequence of the protein is shown as SEQ ID NO. 2. Wherein the secondary structure of the protein is mainly irregular curl and alpha-helix, and is stable protein, and the number of amino acids of the protein is 601. In a third aspect, the present invention also provides the use of a recombinant expression vector, a transient expression vector comprising gene PbTCP, in (A1) - (A3) as follows: (A1) Application of reducing lignin content in plants; (A2) Use in the preparation of a product for reducing lignin content in plants; (A3) Use in breeding to reduce lignin content in plants. The present invention can construct recombinant expression vector containing the gene PbTCP with available plant expression vector. When the gene PbTCP is used for constructing a recombinant plant over-expression vector, a cauliflower mosaic virus (CAMV) 35S strong promoter can be added before the transcription initiation nucleotide, and when the gene of the invention is used for constructing a plant expression vector, ATG can be used as an initiation codon, but the ATG must be the same as the reading frame of a coding sequence so as to ensure the correct translation of the whole sequence. To facilitate the identification and selection of transgenic plants, the plant expression vectors used are processed, and genes encoding compounds which produce luminescence (luciferase genes), antibiotic markers with resistance (kanamycin markers) expressed in plants are added. From the safety aspect of transgenic plants, the transformed plants can be directly screened by hygromycin without adding any selectable marker gene. In a specific embodiment, the backbone vector of the recombinant expression vector is pCAMBIA1300-GFP. In a fourth aspect, the present invention also provides the use of a recombinant bacterium comprising the gene PbTCP as described above in (A1) to (A3) as follows: (A1) Application of reducing lignin content in plants; (A2) Use in the preparation of