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CN-122012553-A - Rice chalkiness related gene Chalk3, encoding protein and application thereof

CN122012553ACN 122012553 ACN122012553 ACN 122012553ACN-122012553-A

Abstract

The invention discloses a rice grain chalky related gene Chalk3, a coded protein and application thereof, wherein the gene Chalk3 provided by the invention is 1) a DNA molecule shown as SEQ ID NO.1, 2) a DNA molecule shown as SEQ ID NO.2, 3) a DNA molecule which hybridizes with a DNA sequence limited by 1) or 2) under strict conditions and codes the protein, and 4) a DNA molecule which has more than 90% homology with the DNA sequence limited by 1) or 2) or 3) and codes rice grain chalky Bai Xiangguan protein. The invention also provides a protein coded by the gene, which influences the chalkiness degree of rice grains, and the expression level of the Chalk3 gene in the invention is inversely related to the chalkiness degree of the rice grains. Therefore, the Chalk3 gene participates in the chalky regulation of rice grains, and provides a useful gene resource for improving rice quality.

Inventors

  • WAN JIANMIN
  • LIU SHIJIA
  • LIN QIBING
  • ZHU SHANSHAN
  • GUO XIUPING
  • JIANG XIAOKANG
  • REN YULONG
  • WANG YIHUA
  • LEI CAILIN
  • WANG XIN
  • WANG JIE
  • ZHAO ZHICHAO
  • CHENG ZHIJUN

Assignees

  • 中国农业科学院作物科学研究所
  • 南京农业大学

Dates

Publication Date
20260512
Application Date
20251030

Claims (10)

  1. 1. A rice chalky gene Chalk3 is characterized in that the gene is a DNA molecule shown in the following 1) or 2) or 3) or 4): 1) A DNA sequence shown in SEQ ID NO. 1; 2) A DNA sequence shown in SEQ ID NO. 2; 3) A DNA sequence which hybridizes under stringent conditions to the DNA sequence defined in 1) or 2) and which codes for a protein according to SEQ ID NO. 3; 4) A DNA sequence which has more than 90 percent of homology with the DNA sequence defined in 1), 2) or 3) and codes for rice grain chalky character related protein.
  2. 2. A protein encoded by the rice chalky related gene of claim 1.
  3. 3. The protein of claim 2, selected from any one of (a) or (b): (a) A protein consisting of the amino acid sequence shown in SEQ ID NO. 3; (b) And (3) the protein which is obtained by substituting and/or deleting and/or adding one or more amino acid residues in the amino acid sequence of SEQ ID NO.3, is related to the chalky rice grain character and is derived from SEQ ID NO. 3.
  4. 4. A recombinant expression vector, expression cassette, transgenic cell line or recombinant bacterium comprising the gene of claim 1.
  5. 5. The recombinant expression vector according to claim 4, wherein the recombinant expression vector is a recombinant plasmid obtained by inserting the gene according to claim 1 between the multiple cloning sites of pCAMBIA1390 vector.
  6. 6. Primers for amplifying the full length of the gene of claim 1 or any fragment thereof.
  7. 7. The gene as claimed in claim 1, wherein the gene encodes a protein, and the recombinant expression vector, expression cassette, transgenic cell line or recombinant bacterium containing the gene is used for preparing transgenic rice with different chalkiness or improving rice quality of rice, specifically, the chalkiness gene chalks 3 of rice is edited or knocked out or overexpressed in a target rice material, and the expression level of the chalks 3 gene is changed, so that rice plants with different chalks phenotypes are obtained, and the rice quality is improved.
  8. 8. The use according to claim 7, wherein the use is to edit or knock out the chalky 3 gene of rice, such that the expression level of the chalky 3 gene in the rice variety of interest is altered, thereby obtaining rice plants with different chalky phenotypes; the gene knockout employs a CRISPR/Cas9 system.
  9. 9. The use according to claim 7, characterized in that the starting rice has plants with abnormal endosperm development due to the defect of the gene indicated by SEQ ID No.2, i.e. an abnormal increase in rice chalkiness; The application is that the gene as set forth in claim 1 is introduced into initial rice with abnormal endosperm development caused by SEQ ID NO.2 gene defect to obtain transgenic rice with normal endosperm development or obtain transgenic rice with improved rice quality.
  10. 10. A method for breeding a transgenic plant having a normal endosperm, which comprises introducing the gene of claim 1 into a plant having a endosperm dysplasia caused by a deficiency in the gene of SEQ ID No.2, to obtain a transgenic plant having a normal endosperm, specifically, introducing the gene into a plant having a endosperm dysplasia by the recombinant expression vector of claim 4 or 5.

Description

Rice chalkiness related gene Chalk3, encoding protein and application thereof Technical Field The invention belongs to the field of genetic engineering, and particularly relates to a rice chalky related gene Chalk3, and a coding protein and application thereof. Background Rice is one of important grain crops in China, and the rice yield realizes qualitative leap through multiple green revolution. With the continuous improvement of living standard, the consumption demand of people for rice is gradually changed from 'full' to 'good eating and healthy eating', and the market demand of high-quality rice is continuously increased. Appearance quality is the most intuitive index for measuring rice quality, wherein chalkiness are key factors for limiting rice quality and market value, not only influence appearance, but also influence taste quality to a certain extent, and the commodity value is directly determined by the level of chalkiness. In order to meet the demands of consumers on high-quality rice, the rapid improvement of chalky characters and cultivation of varieties with good appearance become urgent tasks in the field of rice breeding, and have important significance in improving the quality and yield of rice. Meanwhile, with the development of special rice breeding, special products such as sake brewed rice and low gluten rice are also increasingly paid attention. Unlike edible rice, chalky has a positive effect in sake brewing rice, and provides a new direction for rice quality improvement and diversified breeding. Chalky refers to the opaque, loose white portion of the rice endosperm, which can be divided into heart, stomach and back, with the highest incidence of stomach whitening. Chalkiness (DEC) and chalkiness (PGWC) are the main measures of chalkiness. Chalkiness are complex quantitative traits, cooperatively controlled by multiple genes, and significantly affected by genetic background and environmental conditions. In recent years, scholars at home and abroad have made a certain progress in chalky character research, and the occurrence mechanism of the scholars is deeply studied from the aspects of QTL detection and positioning, gene expression regulation and control, related protein activity, cultivation conditions and the like. However, most studies have focused mainly on chalky physiological regulation and its correlation with other traits, and the analysis of their genetic basis has yet to be enhanced. The formation of chalky phenotype involves multiple genetic regulation pathways, mainly including multiple links of starch synthesis, protein synthesis and transport, transcription regulatory factors, and organelle development, which directly or indirectly affect starch and protein accumulation. Therefore, the novel chalky regulation gene is further mined and cloned, the molecular action mechanism of the chalky regulation gene is clarified and applied to breeding practice, and the novel chalky regulation gene has important theoretical significance and application value for promoting rice quality improvement and molecular breeding. Disclosure of Invention The invention aims to disclose a rice chalky related gene Chalk3, and a coding protein and application thereof. The invention provides a rice chalky 3 regulatory gene, wherein the chalky 3 gene is positioned on a No.9 chromosome of rice, and the gene number is Os03g0248600 (RAP-DB naming rule) or C (MSU naming rule). The gene Chalk3 provided by the invention is a DNA molecule as described in the following 1) or 2) or 3) or 4): 1) A DNA molecule shown in SEQ ID No. 1; 2) A DNA molecule shown in SEQ ID No. 2; 3) A DNA molecule which hybridizes under stringent conditions to the DNA sequence defined in 1) or 2) and which encodes said protein; 4) A DNA molecule which has more than 90% homology with the DNA sequence defined in 1) or 2) or 3) and which codes for rice grain chalky Bai Xiangguan protein. The invention also provides a protein coded by the gene Chalk 3. Specifically, the protein provided by the invention is selected from any one of the following (a) or (b): (a) A protein consisting of the amino acid sequence shown in SEQ ID NO. 3; (b) And (3) a protein which is related to rice grain chalkiness and is derived from the SEQ ID NO.3 through substitution and/or deletion and/or addition of one or more amino acid residues of the amino acid sequence of the SEQ ID NO. 3. The invention also provides a recombinant expression vector, an expression cassette, a transgenic cell line or recombinant bacteria containing the gene Chalk 3. Recombinant expression vectors containing any of the above genes are also within the scope of the present invention. Recombinant expression vectors containing the genes can be constructed using existing plant expression vectors. The plant expression vector comprises a binary agrobacterium vector, a vector which can be used for plant microprojectile bombardment and the like. The plant expression vector may also comprise the