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CN-121975835-A - Application of MSP gene in regulating and controlling grain number per ear of rice and application

CN121975835ACN 121975835 ACN121975835 ACN 121975835ACN-121975835-A

Abstract

The invention discloses application of MSP gene to regulation of rice grain number per spike, wherein the CDS sequence of the MSP gene is shown as SEQ ID No.1, the amino acid sequence of encoding protein is shown as SEQ ID No.2, and the MSP gene is used as a transcription inhibitor and co-inhibitor OsTPR2 to interact, specifically bind with the P4 locus containing CArG-box of the downstream target gene TAWAWA1 promoter to inhibit TAWAWA1 expression, regulate rice inflorescence morphology and regulate grain number per spike. The invention also utilizes the promoter of the glume-protecting/glume-specific expression gene to be fused with the MSP gene CDS to construct a fusion vector, converts MSP mutants, obtains incompletely complementary plants which are normal in glume-protecting, recover in grain size and fruiting rate and have the grain number per spike still higher than that of wild plants, and solves the defects of small grains and the like of the MSP mutants. The invention expands the rice inflorescence development regulation theory and provides a theoretical basis and a new thought for high-yield molecular breeding.

Inventors

  • LI YUNFENG
  • ZHANG CHANGWEI
  • YANG ZHENGLIN
  • HE GUANGHUA
  • ZHUANG HUI
  • CHEN HAO
  • ZHANG TING
  • LING YINGHUA
  • ZHU XIAOYAN
  • SHEN WENQIANG
  • SANG XIANCHUN
  • WANG NAN

Assignees

  • 西南大学
  • 宜宾西南大学研究院

Dates

Publication Date
20260505
Application Date
20251016

Claims (10)

  1. 1. The application of MSP genes or MSP gene coding proteins in regulating and controlling the grain number per spike of rice is characterized in that a CDS sequence of the MSP genes is shown as SEQ ID No.1, an amino acid sequence of the MSP gene coding proteins is shown as SEQ ID No.2, the application is realized by adopting MSP as a transcription inhibitor, interacting with a co-inhibitor OsTPR2, specifically combining with a promoter region of a downstream target gene TAWAWA1 and recruiting histone deacetylase to inhibit the expression of TAWAWA1 genes, wherein the promoter region is a P4 site containing CArG-box, and a nucleotide sequence of the P4 site is shown as SEQ ID No. 3.
  2. 2. A method for regulating the grain number per ear of rice is characterized by comprising the steps of regulating the expression of a rice TAWAWA gene by using an MSP gene or an MSP gene coding protein, regulating the grain number per ear of rice, wherein the CDS sequence of the MSP gene is shown as SEQ ID No.1, and the amino acid sequence of the MSP gene coding protein is shown as SEQ ID No. 2.
  3. 3. The method of claim 2, comprising reducing or silencing expression of the MSP gene by gene-induced mutation techniques, gene knockout techniques, gene interference expression techniques, over-expressing the downstream target gene TAWAWA1, and increasing the number of grains per spike in rice.
  4. 4. The application of the MSP gene or the MSP gene coding protein in rice yield improvement breeding is characterized in that the CDS sequence of the MSP gene is shown as SEQ ID No.1, the amino acid sequence of the MSP gene coding protein is shown as SEQ ID No.2, and the application is that the expression of rice TAWAWA gene is regulated by the MSP gene or the MSP gene coding protein, and the grain number per spike of rice is regulated.
  5. 5. A method for improving rice yield is characterized by comprising the steps of constructing a fusion vector by utilizing a promoter of a glume-protecting/glume-specific expression gene and CDS of an MSP gene, transforming MSP mutant to obtain an incomplete complementary plant with normal glume protection, recovered grain size and seed setting rate and still increased grain number per ear compared with wild type, wherein the MSP mutant is a maintainer indica rice large 1B mutant, the phenotype is that the number of branches and stems and the number of spikelets are obviously increased, but grains are small, glume protection is elongated, and the CDS sequence of the MSP mutant gene is shown as SEQ ID No. 4.
  6. 6. The method of claim 5, wherein the glume-specific expression gene is G1, NSG1, or OsbZIP26, the promoter sequence of G1 is shown in SEQ ID No.5, the promoter sequence of NSG1 is shown in SEQ ID No.6, and the promoter sequence of OsbZIP26 is shown in SEQ ID No. 7.
  7. 7. A rice yield improvement breeding method, characterized in that the method comprises the following steps: (1) Constructing a fusion vector by fusing a promoter of a glume-protecting/glume-specific expression gene with CDS of an MSP gene, wherein the CDS sequence of the MSP gene is shown as SEQ ID No. 1; (2) Transforming MSP mutant, namely transforming MSP mutant through agrobacterium mediation by the fusion vector in the step (1), obtaining an incomplete complementary plant, wherein the glume-protecting morphology of the incomplete complementary plant is normal, the grain size and the fruiting rate are recovered, the grain number per spike is increased compared with that of wild type, the MSP mutant is a maintainer line indica rice large 1B mutant, the phenotype is that the branch number and the spike number are obviously increased, the grain is small, the glume-protecting elongation is achieved, and the MSP gene mutation CDS sequence is shown as SEQ ID No. 4.
  8. 8. The method of claim 7, wherein the glume-specific expression gene is G1, NSG1, or OsbZIP26, the promoter sequence of G1 is shown in SEQ ID No.5, the promoter sequence of NSG1 is shown in SEQ ID No.6, and the promoter sequence of OsbZIP26 is shown in SEQ ID No. 7.
  9. 9. The fusion vector for rice yield improvement breeding is characterized by being constructed by fusing a promoter of a glume protection/glume specific expression gene and a CDS of an MSP gene, wherein the sequence of the CDS of the MSP gene is shown as SEQ ID No. 1.
  10. 10. The fusion vector of claim 9, wherein the glume-protecting/glume-specific expression gene is G1, NSG1, or OsbZIP26, the promoter sequence of G1 is shown in SEQ ID No.5, the promoter sequence of NSG1 is shown in SEQ ID No.6, and the promoter sequence of OsbZIP26 is shown in SEQ ID No. 7.

Description

Application of MSP gene in regulating and controlling grain number per ear of rice and application Technical Field The invention relates to the technical field of plant genetic engineering and rice breeding, in particular to application of MSP genes to regulate and control the grain number per ear of rice and application thereof. Background Rice is also an important food crop as a monocotyledonous mode plant. The three elements of the rice yield are the number of spikes per mu, the number of grains per spike and thousand grain weight. Wherein the grain number per ear is one of key factors for the yield of gramineous grain crops such as rice and the like. In recent years, the regulatory mechanism of inflorescence structure has been greatly advanced, and a plurality of genes related to rice spike development are identified, and the genes positively or negatively regulate various spike morphological characters, so as to determine the number of grains per spike, and finally play an important role in the rice yield constitution. In previous studies scientists have identified and cloned genes that are partially related to the number of grains per ear. Such as LAX1, MOC1, RCN2, APO1, APO2 and TAWAWA1 (TAW 1 for short), and the like, which increase the number of grains per spike by regulating morphological characters of the spike, such as the number of branches, the length, the seed attachment density, and the like, thereby affecting the yield. In summary, genes affecting the number of grains per ear are involved in multiple regulatory pathways and are enormous in number. However, there are relatively few genes actually used in production to improve yield, mainly including SP1, DEP2/EP2, osSPL/IPA 1, gn1a/OsCKX2 and TAWAWA (TAW 1 for short), etc. The main reason is that many gene functions have multiple effects, and the number of grains per ear is increased after mutation, but other adverse properties such as reduced tillering, overlong growth period, reduced fruiting rate, reduced thousand seed weight and the like are brought at the same time. Therefore, the current rice yield breeding based on the grain number per ear should be cut in from two aspects, namely, developing new gene resources for regulating the grain number per ear, which still has the primary significance, and carrying out molecular design breeding on related genes, regardless of natural mutants or simple transgenes, and reducing or eliminating the influence of adverse traits while retaining or even exerting the beneficial traits by proper molecular design on the basis of analyzing the molecular mechanisms of the genes. Disclosure of Invention The invention aims to provide an application of MSP genes in regulating and controlling the grain number per ear of rice and an application thereof in improving the yield of the rice. In order to achieve the above object, the present invention provides the following technical solutions: The invention provides an application of MSP genes or MSP gene coding proteins in regulating and controlling the grain number per spike of rice, wherein the CDS sequence of the MSP genes is shown as SEQ ID No.1, the amino acid sequence of the MSP gene coding proteins is shown as SEQ ID No.2, the application is realized by adopting MSP as a transcription inhibitor, interacting with a co-inhibitor OsTPR2, specifically combining with a promoter region of a downstream target gene TAWAWA1 and recruiting histone deacetylase to inhibit the expression of TAWAWA1 genes, the promoter region is a P4 site containing CArG-box, and the nucleotide sequence of the P4 site is shown as SEQ ID No. 3. The invention also provides a method for regulating the grain number per ear of rice, which comprises regulating the expression of rice TAWAWA gene by using MSP gene or MSP gene coding protein, regulating the grain number per ear of rice, wherein the CDS sequence of the MSP gene is shown as SEQ ID No.1, and the amino acid sequence of the MSP gene coding protein is shown as SEQ ID No. 2. Further, the method comprises the steps of reducing or silencing the expression of the MSP gene through a gene induction mutation technology, a gene knockout technology and a gene interference expression technology, so that the downstream target gene TAWAWA gene is over-expressed, and the grain number per spike of rice is improved. The invention also provides application of the MSP gene or the MSP gene coding protein in rice yield improvement breeding, wherein the CDS sequence of the MSP gene is shown as SEQ ID No.1, the amino acid sequence of the MSP gene coding protein is shown as SEQ ID No.2, and the application is that the expression of rice TAWAWA gene is regulated and the grain number per spike of rice is regulated and controlled through the MSP gene or the MSP gene coding protein. The invention also provides a method for improving rice yield, which comprises the steps of constructing a fusion vector by utilizing a promoter of a glume-protecting/glume-specific