CN-121992003-A - Soybean GmSMS protein and application of encoding gene thereof in regulating quality traits of soybean seeds

Abstract

The invention provides application of soybean GmSMS protein and a coding gene thereof in regulating and controlling quality traits of soybean seeds. GmSMS6 genome sequence is shown as SEQ ID No.1, and encoded amino acid sequence is shown as SEQ ID No. 2. According to the invention, soybean Zp661 is subjected to gene editing through CRSIPR-Cas9 system, so that homozygous mutant plants Z1 and Z2 with obviously increased seed protein content and obviously reduced oil content compared with wild soybean Zp661 are obtained, and the function of soybean GmSMS in the aspect of regulating and controlling soybean grain quality is proved. The invention provides important gene resources and theoretical guidance for soybean molecular breeding and germplasm innovation.

Inventors

• HE CHAOYING
• LI BINGBING
• WANG YAN

Assignees

• 中国科学院植物研究所

Dates

Publication Date

20260508

Application Date

20241106

Claims (10)

1. 1. A method of modulating a plant seed trait, the method comprising modulating a plant seed trait of interest by gene knockout of a gene encoding a target protein in a plant of interest, the plant of interest comprising a gene encoding the target protein; The target protein is GmSMS, and is any one of the following: A1 Amino acid sequence is a protein shown in sequence 2, A2 Protein which is obtained by substituting and/or deleting and/or adding amino acid residues of the protein A1), has more than 80 percent of identity with the protein A1) and can regulate the properties of plant seeds, A3 Fusion proteins obtained by ligating the N-terminal or/and C-terminal of A1) or A2) with a protein tag.
2. 2. A method of growing a plant with altered seed grain properties comprising obtaining a plant with altered seed grain properties of a plant of interest by gene knockout, said plant of interest comprising a gene encoding a protein of claim 1.
3. 3. The method according to claim 1 or 2, wherein the protein is derived from soybean.
4. 4. A method according to claims 1-3, wherein the genome of the plant of interest comprises a DNA molecule having a nucleotide sequence that is sequence 1 of the sequence listing, and wherein the gene knockout comprises any of the following procedures for the genome of the plant of interest: B1 Deletion of 342 th-344 th nucleotides of sequence 1 in the sequence table of the target plant, and insertion of an adenine ribonucleotide between 491 th and 492 th nucleotides; b2 The 336 th-340 th nucleotide of the sequence 1 in the sequence table of the target plant is deleted.
5. 5. The method of any one of claims 1-4, wherein the plant grain trait is increased plant grain protein content and/or decreased plant grain oil content.
6. 6. The method of any one of claims 1-5, wherein the plant is any one of: C1 Dicotyledonous plants, C2 A plant of the order Rosales, C3 A) a leguminous plant, C4 A plant of the genus glycine, C5 Soy bean).
7. 7. Use of a biomaterial, characterized in that the use is any one of the following: d1 The application of the biological material in regulating and controlling the plant grain character and/or preparing a product for regulating and controlling the target plant grain character, D2 Use of said biological material in plant breeding and/or in the preparation of plant breeding products; the biological material is protein, a substance for regulating and controlling the expression of the encoding gene of the protein or a substance for regulating and controlling the content of the protein; The protein is GmSMS and is any one of the following: e1 Amino acid sequence is a protein shown in sequence 2, E2 Protein which is obtained by substituting and/or deleting and/or adding amino acid residues of the protein E1), has more than 80 percent of identity with the protein E1) and can regulate the properties of plant seeds, E3 Fusion proteins obtained by ligating the N-terminal or/and C-terminal of E1) or E2) with a protein tag.
8. 8. The use according to claim 7, wherein the substance is any one of the following: F1 A nucleic acid molecule which inhibits or reduces or down-regulates the expression of a gene encoding a protein according to claim 7, F2 Expression of the gene encoding the nucleic acid molecule of F1), F3 An expression cassette containing the gene described under F2), F4 A recombinant vector containing the gene of F2) or a recombinant vector containing the expression cassette of F3), F5 A recombinant microorganism containing the gene of F2), a recombinant microorganism containing the expression cassette of F3), or a recombinant microorganism containing the recombinant vector of F4), F6 A transgenic plant cell line containing F2) said gene, or a transgenic plant cell line containing F3) said expression cassette, or a transgenic plant cell line containing F4) said recombinant vector, F7 A transgenic plant tissue containing F2) said gene, or a transgenic plant tissue containing F3) said expression cassette, or a transgenic plant tissue containing F4) said recombinant vector, F8 A transgenic plant organ containing the gene of F2), or a transgenic plant organ containing the expression cassette of F3), or a transgenic plant organ containing the recombinant vector of F4).
9. 9. The use according to claim 7 or 8, wherein the modulation of plant grain traits is an increase in plant grain protein content and/or a decrease in plant grain oil content.
10. 10. The use according to any one of claims 7 to 9, wherein the plant is any one of the following: G1 Dicotyledonous plants, G2 A plant of the order Rosales, G3 A) a leguminous plant, G4 A plant of the genus glycine, G5 Soy bean).

Description

Soybean GmSMS protein and application of encoding gene thereof in regulating quality traits of soybean seeds Technical Field The invention belongs to the field of variation or genetic engineering, and particularly relates to application of soybean GmSMS protein and a coding gene thereof in regulating and controlling quality traits of soybean seeds. Background Soybean (Glycine max) is an important dual-purpose crop for food and feed, and provides rich grease, protein and other nutrient substances for human beings and animals. Although China is the origin of soybeans, soybean breeding has been slow in recent years, and the yield per unit area is relatively backward compared with the world major soybean production countries. At present, the domestic soybean has serious insufficient self-sufficiency rate, higher external dependence and insufficient self-sufficiency rate of 20 percent. The soybean seed protein and oil content is a polygenic controlled quantitative trait and is also affected by environmental factors. The protein and oil content of soybean seeds are determined by multiple factors and multiple regulation ways, and although some researches report on gene loci for regulating the oil content of soybean proteins at present, the functional identification and regulation network of key genes are not clear. The molecular mechanism formed by the soybean seed protein and the oil content is analyzed, a cooperative regulation network between the soybean seed protein and the oil content is revealed, and the method is an important theoretical basis for creating high-yield and high-quality soybean varieties. In recent years, with the continuous and deep research of soybean genomics and modern molecular biology, the protein and grease content of soybean seeds can be accurately regulated and controlled by means of gene orientation improvement and the like, so as to achieve the breeding goal of high-yield and high-quality soybean varieties. Disclosure of Invention The invention aims to solve the technical problem of regulating and controlling the quality of plants, especially soybean seeds by using biological materials. To this end, the invention provides a method for regulating and controlling plant seed traits, which is characterized in that the method comprises regulating and controlling target plant seed traits by gene knockout, wherein the gene knockout is used for knocking out a target protein coding gene in target plants, and the target plants contain the target protein coding gene; The target protein is GmSMS, and is any one of the following: A1 Amino acid sequence is a protein shown in sequence 2, A2 Protein which is obtained by substituting and/or deleting and/or adding amino acid residues of the protein A1), has more than 80 percent of identity with the protein A1) and can regulate the properties of plant seeds, A3 Fusion proteins obtained by ligating the N-terminal or/and C-terminal of A1) or A2) with a protein tag. The invention also provides a method for cultivating the plant with changed seed grain shape, which comprises the step of obtaining the plant with changed seed grain shape of the target plant through gene knockout, wherein the target plant contains the encoding gene of the protein. The protein is derived from soybean. In the above method, the genome of the target plant contains a DNA molecule with a nucleotide sequence of sequence 1 in a sequence table, and the gene knockout comprises any one of the following operations on the genome of the target plant: B1 Deletion of 342 th-344 th nucleotides of sequence 1 in the sequence table of the target plant, and insertion of an adenine ribonucleotide between 491 and 492 th nucleotides. B2 The 336 th-340 th nucleotide of the sequence 1 in the sequence table of the target plant is deleted. The gene knockout refers to a phenomenon in which a specific target gene is inactivated by homologous recombination. Gene knockout is the inactivation of a particular target gene by a change in DNA sequence. The target sequence for the gene knockout may be sgRNA1:5'-GTCGCCTACAAGAACGTAAT-3' and/or sgRNA2:5'-CAAGCTCCTCGACACGCGCC-3'. The gene knockout comprises the step of introducing a gene knockout vector taking 5'-GTCGCCTACAAGAACGTAAT-3' and/or sgRNA2:5'-CAAGCTCCTCGACACGCGCC-3' as a target sequence into the target plant. The invention also protects the application of the biological material, wherein the application is any one of the following: C1 The application of the biological material in regulating and controlling the plant grain character and/or preparing a product for regulating and controlling the target plant grain character, C2 Use of said biological material in plant breeding and/or in the preparation of plant breeding products; the biological material is protein, a substance for regulating and controlling the expression of the encoding gene of the protein or a substance for regulating and controlling the content of the protein; The protein is GmSMS and is