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CN-121992033-A - Application of alfalfa cFBP protein in regulation and control of plant types

CN121992033ACN 121992033 ACN121992033 ACN 121992033ACN-121992033-A

Abstract

The application belongs to the field of plant genetic engineering, and provides application of alfalfa cFBP protein in regulation of plant types, wherein the nucleotide sequence of a alfalfa cFBP protein coding gene is SEQ ID NO.1 or SEQ ID NO.3. The alfalfa cFBP protein coding gene positively regulates the plant height, the branch number and the biomass of alfalfa, and the plant height, the branch number and the biomass of the plant after MscFBP gene is over-expressed are obviously improved compared with those of a control material. The application successfully obtains the transgenic alfalfa with obviously improved biological yield by over-expressing MscFBP genes through the genetic engineering technology, and provides important paradigms and gene resources for cultivating high-yield plant varieties.

Inventors

  • WANG TAO
  • Ye Qinyi
  • DONG JIANGLI
  • GAO YAJUAN

Assignees

  • 中国农业大学

Dates

Publication Date
20260508
Application Date
20260402

Claims (10)

  1. 1. The application of the alfalfa cFBP protein in regulating plant types is characterized in that the amino acid sequence of the alfalfa cFBP protein is SEQ ID NO.2 or SEQ ID NO.4.
  2. 2. The use according to claim 1, wherein the expression level of alfalfa cFBP protein or its coding gene in the plant is increased to achieve one or more effects selected from (A1) - (A3): (A1) The plant height is improved; (A2) Increasing the number of plant branches; (A3) Improving plant biomass.
  3. 3. The use according to claim 1 or 2, wherein increasing the expression level of alfalfa cFBP protein or its coding gene in said plant is achieved by one or more means selected from (B1) - (B2): (B1) Means for overexpressing the gene encoding alfalfa cFBP protein; (B2) Means for increasing the activity of alfalfa cFBP proteins.
  4. 4. The use according to claim 3, (B1) is a nucleic acid molecule encoding a gene of alfalfa cFBP protein, or a vector, expression cassette, recombinant microorganism, plant cell, plant tissue or plant organ comprising a gene encoding a alfalfa cFBP protein, preferably an overexpressing vector comprising a gene encoding a alfalfa cFBP protein, more preferably the overexpressing vector comprises a constitutive promoter, still more preferably a pCAMBIA1307 vector comprising a gene encoding a alfalfa cFBP protein.
  5. 5. The use according to any one of claims 1-4, wherein the plant is alfalfa.
  6. 6. The use according to any one of claims 1-5 for one or more of (C1) - (C3): (C1) Improving plant type; (C2) Providing plant propagation material, preferably seeds, tissue culture material, organs available for propagation, of improved plant type; (C3) Plant lines of improved plant type are provided.
  7. 7. The alfalfa cFBP protein is characterized in that the amino acid sequence of the alfalfa cFBP protein is SEQ ID NO.2 or SEQ ID NO.4.
  8. 8. The alfalfa cFBP protein-encoding gene according to claim 7.
  9. 9. The coding gene according to claim 8, wherein the nucleotide sequence of the coding gene is SEQ ID NO.1 or SEQ ID NO.3.
  10. 10. Use of any one of the means (D1) - (D3) for detecting the expression level of alfalfa cFBP protein and the expression level of the alfalfa cFBP protein-encoding gene: (D1) Judging the plant type of alfalfa plants; (D2) Predicting plant types of alfalfa propagation materials; (D3) Cultivating alfalfa varieties with improved plant types; Means for detecting the expression level of alfalfa cFBP protein and the expression level of alfalfa cFBP protein-encoding gene are one or more of the following: (E1) A primer; (E2) A probe; (E3) A PCR reagent; (E4) A chip; (E5) An antibody; (E6) ELISA reagents; The nucleotide sequence of the alfalfa cFBP protein coding gene is SEQ ID NO.1 or SEQ ID NO.3, and the amino acid sequence of the alfalfa cFBP protein is SEQ ID NO.2 or SEQ ID NO.4.

Description

Application of alfalfa cFBP protein in regulation and control of plant types Technical Field The application belongs to the field of plant genetic engineering, and particularly provides application of alfalfa cFBP protein in regulation of plant types. Background The plant type is an important agronomic character for determining the crop yield, and comprises a plurality of aspects such as plant height, branch/tillering number, branch/tillering angle and the like, and the yield formation is directly influenced by regulating and controlling the utilization efficiency of plants on resources such as light, water, fertilizer and the like. The ideal plant type concept is proposed for the first time in 1968, and refers to a form in which the competition intensity among individual crops is minimum and the accumulation of grain assimilation substances is maximum, namely the greatest common divisor capable of efficiently utilizing light energy and land fertility and achieving crop yield and quality. The ideal plant type has become an important breeding target of crops such as rice, corn, wheat, soybean, rape and the like, but a clear concept is not formed in most forage grass, so that the excavation of genes related to plant type regulation and control of the crops and the elucidation of molecular mechanisms thereof are of great significance for obtaining a variety with higher yield. Alfalfa (Medicago sativa l.) is known as "grass king", and is widely planted worldwide and is a representative of perennial quality forage grass. Plant type is a key factor influencing the yield of alfalfa, and cultivation of alfalfa varieties with upright plant type, rapid growth and large biomass is an urgent industrial demand. Disclosure of Invention In one aspect, the application provides application of alfalfa cFBP protein in regulating plant types, and the amino acid sequence of the alfalfa cFBP protein is SEQ ID NO.2 or SEQ ID NO.4. Further, the use increases the expression level of alfalfa cFBP protein or its encoding gene in the plant to achieve one or more effects selected from (A1) - (A3); (A1) The plant height is improved; (A2) Increasing the branch number of plants; (A3) Improving plant biomass. Further, the expression level of alfalfa cFBP protein or its coding gene in the plant is achieved by one or more means selected from (B1) - (B2): (B1) Means for overexpressing the gene encoding alfalfa cFBP protein; (B2) Means for increasing the activity of alfalfa cFBP proteins. Further, (B1) is a nucleic acid molecule encoding a gene of alfalfa cFBP protein, or a vector, an expression cassette, a recombinant microorganism, a plant cell, a plant tissue or a plant organ comprising a gene encoding a alfalfa cFBP protein, preferably an over-expression vector comprising a gene encoding a alfalfa cFBP protein, more preferably the over-expression vector comprises a constitutive promoter, still more preferably a pCAMBIA1307 vector comprising a gene encoding a alfalfa cFBP protein; furthermore, known techniques such as CRISPRa activation or stimulation of expression by saRNA may be used. Further, the plant is alfalfa. Further, the application is for one or more of (C1) - (C3): (C1) Improving plant type; (C2) Providing plant propagation material, preferably seeds, tissue culture material, organs available for propagation, of improved plant type; (C3) Plant lines of improved plant type are provided. In another aspect, the present application provides alfalfa cFBP protein, the amino acid sequence of which alfalfa cFBP protein is SEQ ID No.2 or SEQ ID No.4. In another aspect, the application provides a coding gene of the alfalfa cFBP protein. Further, the nucleotide sequence of the coding gene is SEQ ID NO.1 or SEQ ID NO.3. In another aspect, the present application provides the use of any one of the means (D1) - (D3) for detecting the expression level of alfalfa cFBP protein and the expression level of the gene encoding alfalfa cFBP protein: (D1) Judging the plant type of alfalfa plants; (D2) Predicting plant types of alfalfa propagation materials; (D3) Cultivating alfalfa varieties with improved plant types. Means for detecting the expression level of alfalfa cFBP protein and the expression level of alfalfa cFBP protein-encoding gene are one or more of the following: (E1) A primer; (E2) A probe; (E3) A PCR reagent; (E4) A chip; (E5) An antibody; (E6) ELISA reagents. The nucleotide sequence of the alfalfa cFBP protein coding gene is SEQ ID NO.1 or SEQ ID NO.3, and the amino acid sequence of the alfalfa cFBP protein is SEQ ID NO.2 or SEQ ID NO.4. The codons of the protein may be varied by those skilled in the art according to the requirements of the host or the needs of the expression, according to known codon preferences, and the like. Two MscFBP gene haplotypes MscFBP Hap1 and MscFBP Hap2 for positively regulating alfalfa plant height, branch number and biomass are discovered. By knocking MscFBP out the gene editing technology, the plant heigh