Search

CN-122012608-A - Gene PbMYB20 for regulating and controlling pear fruit stone cell formation

CN122012608ACN 122012608 ACN122012608 ACN 122012608ACN-122012608-A

Abstract

The invention discloses a gene PbMYB20 for regulating and controlling pear fruit lignin synthesis, 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 fruit of the Dangshan pear, the content of stone cells and lignin is obviously increased compared with the injection of no-load control parts, the gene construction silencing vector is injected into the fruit of the Dangshan pear, the content of stone cells and lignin is obviously reduced compared with the injection of no-load control parts, the gene construction over-expression vector is introduced into the Arabidopsis, the secondary cell walls of the duct cells of the obtained PbMYB20 transgenic Arabidopsis are obviously thickened, and the accumulation of lignin in stems is obviously increased. 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
  • TIAN JIA
  • QI KAIJIE
  • XIE ZHIHUA

Assignees

  • 南京农业大学三亚研究院
  • 南京农业大学
  • 新疆农业大学

Dates

Publication Date
20260512
Application Date
20260416

Claims (10)

  1. 1. Use of a substance that reduces the protein content or activity of PbMYB20, or reduces/silences the expression level of the gene PbMYB20, which is any of the following, for reducing the lignin content in pear fruits or arabidopsis: a1 Nucleic acid molecules that silence the expression level of the gene PbMYB 20; a2 An expression cassette comprising a 1) said nucleic acid molecule; a3 A recombinant vector comprising a nucleic acid molecule according to a 1) or a recombinant vector comprising an expression cassette according to a 2); a4 A) a recombinant microorganism comprising a 1) said nucleic acid molecule, or a recombinant microorganism comprising a 2) said expression cassette, or a recombinant microorganism comprising a 3) said recombinant vector; Wherein the CDS sequence of the gene PbMYB20 is shown in SEQ ID NO. 1.
  2. 2. Use of a substance that reduces the protein content or activity of PbMYB20, or reduces/silences the expression level of the gene PbMYB20, in the preparation of a product that reduces the lignin content in pear fruits or arabidopsis, said substance that reduces/silences the expression level of the gene PbMYB20 being any of the following: a1 Nucleic acid molecules that silence the expression level of the gene PbMYB 20; a2 An expression cassette comprising a 1) said nucleic acid molecule; a3 A recombinant vector comprising a nucleic acid molecule according to a 1) or a recombinant vector comprising an expression cassette according to a 2); a4 A) a recombinant microorganism comprising a 1) said nucleic acid molecule, or a recombinant microorganism comprising a 2) said expression cassette, or a recombinant microorganism comprising a 3) said recombinant vector; Wherein the CDS sequence of the gene PbMYB20 is shown in SEQ ID NO. 1.
  3. 3. Use of a substance that reduces the protein content or activity of PbMYB20, or reduces/silences the expression level of the gene PbMYB20, in breeding for reducing the lignin content in pear fruits or arabidopsis, said substance that reduces/silences the expression level of the gene PbMYB20 being any of the following: a1 Nucleic acid molecules that silence the expression level of the gene PbMYB 20; a2 An expression cassette comprising a 1) said nucleic acid molecule; a3 A recombinant vector comprising a nucleic acid molecule according to a 1) or a recombinant vector comprising an expression cassette according to a 2); a4 A) a recombinant microorganism comprising a 1) said nucleic acid molecule, or a recombinant microorganism comprising a 2) said expression cassette, or a recombinant microorganism comprising a 3) said recombinant vector; Wherein the CDS sequence of the gene PbMYB20 is shown in SEQ ID NO. 1.
  4. 4. The use according to any one of claims 1 to 3, wherein the backbone vector of the recombinant vector is TRV2.
  5. 5. The method for reducing the lignin content of the pear fruit is characterized by reducing the expression level of a gene PbMYB20 in the pear, wherein the CDS sequence of the gene PbMYB20 is shown as SEQ ID NO. 1.
  6. 6. A method for reducing lignin content in arabidopsis is characterized in that the method is realized by reducing the expression level of a gene PbMYB20 in arabidopsis, and the CDS sequence of the gene PbMYB20 is shown as SEQ ID NO. 1.
  7. 7. A breeding method for reducing the lignin content of pear fruits is characterized in that the method is realized by reducing the expression level of a gene PbMYB20 in pear fruits, and the CDS sequence of the gene PbMYB20 is shown as SEQ ID NO. 1.
  8. 8. A breeding method for reducing lignin content in arabidopsis is characterized by reducing expression quantity of a gene PbMYB20 in arabidopsis, wherein CDS sequence of the gene PbMYB20 is shown as SEQ ID NO. 1.
  9. 9. A method according to any one of claims 5-8, characterized in that the method comprises the steps of: (1) Constructing a recombinant expression vector for silencing the gene PbMYB20 described in claim 1; (2) Transferring the recombinant expression vector constructed in the step (1) into a receptor plant.
  10. 10. The method of claim 9, wherein the backbone vector of the recombinant expression vector is TRV2.

Description

Gene PbMYB20 for regulating and controlling pear fruit stone cell formation Technical Field The invention relates to the fields of plant molecular biology and fruit tree genetic breeding, in particular to a key gene PbMYB20 for regulating and controlling stone cell formation of pear fruits and application of the gene in pear fruit quality improvement. Background Pear is an important fruit tree species in China, pear is deeply favored by consumers by crisp, tender and juicy, sweet and sour and palatable flavor characteristics, and the quality is mainly determined by factors such as peel properties, fruit shape, fruit texture, aroma and the like, wherein stone cells are taken as special thick-wall cells of pear fruits, and are core bottlenecks for limiting fruit texture and commodity value (Jin et al, 2013; kolniak-Ostek, 2016). Studies have shown that stone cell formation is primarily dependent on thickening of secondary cell walls and substantial accumulation of lignin (Tao et al, 2009). When the content of stone cells is too high, the pulp of the pear is rough, the juice is deficient, the hardness is increased, the edible taste of the fruit is seriously reduced, the accumulation of nutrient components such as sugar, organic acid, vitamin and the like in the fruit is influenced, and further, the market price and commodity competitiveness of the fruit are reduced (Choi et al, 2007; zhang et al, 2024). In recent years, researchers have identified key genes that are partially involved in stone cell formation in pear fruits, but since stone cell formation is a complex regulatory process, the inherent complete regulatory mechanism has not been thoroughly elucidated. Based on the method, the key factors for regulating and controlling the stone cell formation of the pear fruit are deeply excavated, and the method has important theoretical support significance and practical production application value for improving the pear fruit quality, promoting the quality improvement and synergy of the pear industry and accelerating the cultivation of high-quality pear varieties. 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, wherein the gene is obtained by separating and cloning a variety 'Dangshan pear' (Pyrus bretschneideri) with high stone cell content, and is named as PbMYB20, the CDS sequence of the gene is shown as SEQ ID NO.1, and the corresponding protein sequence of the gene is shown as a sequence table SEQ ID NO. 2. The discovery of the gene provides new insight for improving the quality of pear fruits. The invention also aims to provide application of the gene PbMYB20 and related biological materials, wherein the gene is constructed into an over-expression vector, the over-expression vector is introduced into arabidopsis through agrobacterium-mediated genetic transformation, and biological function verification shows that the cloned PbMYB20 gene has the functions of promoting lignin accumulation and thickening secondary cell walls, and the lignin content is obviously reduced by transferring the gene constructed silencing vector into pear fruits. The aim of the invention is achieved by the following technical scheme: In a first aspect, the present invention provides the use of a substance that reduces the protein content or activity of PbMYB20, or reduces/silences the expression level of the gene PbMYB20, in reducing the lignin content in pear fruits or Arabidopsis thaliana, said substance that reduces/silences the expression level of the gene PbMYB20 being any of the following: a1 Nucleic acid molecules that silence the expression level of the gene PbMYB 20; a2 An expression cassette comprising a 1) said nucleic acid molecule; a3 A recombinant vector comprising a nucleic acid molecule according to a 1) or a recombinant vector comprising an expression cassette according to a 2); a4 A) a recombinant microorganism comprising a 1) said nucleic acid molecule, or a recombinant microorganism comprising a 2) said expression cassette, or a recombinant microorganism comprising a 3) said recombinant vector; Wherein the CDS sequence of the gene PbMYB20 is shown in SEQ ID NO. 1. In a second aspect, the invention provides the use of a substance that reduces the protein content or activity of PbMYB20, or reduces/silences the expression level of the gene PbMYB20, in the preparation of a product that reduces the lignin content in pear fruits or Arabidopsis thaliana, the substance that reduces/silences the expression level of the gene PbMYB20 being any of the following: a1 Nucleic acid molecules that silence the expression level of the gene PbMYB 20; a2 An expression cassette comprising a 1) said nucleic acid molecule; a3 A recombinant vector comprising a nucleic acid molecule according to a 1) or a recombinant vector comprising an expression cassette according to a 2); a4 A) a recombinant microorganism