Plant-derived glufosinate-resistant glutamine synthetase mutant, nucleic acid molecule and application

A technology of glutamine and nucleic acid molecules, applied in the field of genetic engineering, can solve problems such as interference with plant nitrogen metabolism, insufficient tolerance for commercial application, and impact on plant growth and development, and achieve broad application prospects

Pending Publication Date: 2021-03-30
SICHUAN GEVOTO BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, due to the wave of anti-genetics, the acceptance of genetically modified crops in the world is still low. Even in the Americas with the largest planting area of ​​genetically modified crops, genetically modified crops are mainly limited to several crops such as corn, soybeans, and cotton.
In particular, the bar gene and pat gene are derived from microorganisms, not from the crop itself, which is more likely to cause consumers' resistance
[0007] The glufosinate-ammonium acetylase encoded by Bar gene and pat gene can inactivate glufosinate-ammonium by acetylat

Method used

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  • Plant-derived glufosinate-resistant glutamine synthetase mutant, nucleic acid molecule and application
  • Plant-derived glufosinate-resistant glutamine synthetase mutant, nucleic acid molecule and application
  • Plant-derived glufosinate-resistant glutamine synthetase mutant, nucleic acid molecule and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] The rice (Oryza sativa) glutamine synthetase (GS1) mutant provided in this embodiment is composed of wild-type rice glutamine synthetase itself (named OsGS1_WT, the amino acid sequence is shown in SEQ ID NO.1, and the encoding nucleus The nucleotide sequence is that the 68th amino acid residue S of SEQ ID NO.6) is mutated into A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T , V, W, Y or deleted, and the obtained rice GS1 mutants were named OsA, OsC, OsD, OsE, OsF, OsG, OsH, OsI, OsK, OsL, OsM, OsN, OsP, OsQ, OsR, OsT , OsV, OsW, OsY, and OsX.

[0082] Amino acids of rice GS1 mutants OsA, OsC, OsD, OsE, OsF, OsG, OsH, OsI, OsK, OsL, OsM, OsN, OsP, OsQ, OsR, OsT, OsV, OsW, OsY, OsX and wild-type rice GS1 sequence alignment as figure 1 As shown in the figure: the position indicated by the arrow is the mutation site.

[0083] In this example, the coding sequence of each rice GS1 mutant is at the position encoding the 68th amino acid, and the codons used for the correspon...

Embodiment 2

[0087] The soybean (Glycine max) GS1 mutant provided in this example is derived from the wild-type soybean GS1 itself ((named GmGS1_WT, the amino acid sequence is shown in SEQ ID NO.3, and the encoding nucleotide sequence is SEQ ID NO.8) The 68th position (corresponding to the 68th position of the reference sequence (SEQ ID NO.1)) is mutated from amino acid residue S to D, E, G, H, I, K, M, N, P, Q, V, Y or deleted. The obtained rice soybean GS1 mutants were named GmD, GmE, GmG, GmH, GmI, GmK, GmM, GmN, GmP, GmQ, GmV, GmY and GmX, respectively.

[0088] The amino acid sequence alignment of soybean GS1 mutants GmD, GmE, GmG, GmH, GmI, GmK, GmM, GmN, GmP, GmQ, GmV, GmY, GmX and wild-type soybean GS1 is as follows figure 2 As shown in the figure: the position indicated by the arrow is the mutation site.

[0089] The coding sequence of soybean GS1 mutants GmD, GmE, GmG, GmH, GmI, GmK, GmM, GmN, GmP, GmQ, GmV, GmY and GmX provided in this example corresponds to SEQ ID NO.3.

[0...

Embodiment 3

[0094] The corn (Zea mays) GS1 mutant provided in this example is derived from wild-type corn GS1 itself (named ZmGS1_WT, the amino acid sequence is shown in SEQ ID NO.2, and the encoding nucleotide sequence is SEQ ID NO.7). The 68th position (corresponding to the 68th position of the reference sequence (SEQ ID NO.1)) is mutated from amino acid residue S to A, C, D, E, F, G, H, I, K, L, M, N , P, Q, R, S, T, V, W, Y or deleted. The maize GS1 mutants obtained were named ZmA, ZmC, ZmD, ZmE, ZmF, ZmG, ZmH, ZmI, ZmK, ZmL, ZmM, ZmN, ZmP, ZmQ, ZmR, ZmT, ZmV, ZmW, ZmY and ZmX, respectively.

[0095] Amino acids of maize GS1 mutants ZmA, ZmC, ZmD, ZmE, ZmF, ZmG, ZmH, ZmI, ZmK, ZmL, ZmM, ZmN, ZmP, ZmQ, ZmR, ZmT, ZmV, ZmW, ZmY, ZmX and wild-type maize GS1 sequence alignment as image 3 As shown in the figure: the position indicated by the arrow is the mutation site.

[0096] In this example, the coding sequence of each maize GS1 mutant is at the position encoding the 68th amino acid,...

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Abstract

The invention discloses a plant-derived glutamine synthetase mutant with glufosinate-ammonium resistance, a nucleic acid molecule and application, and relates to the technical field of gene engineering. Compared with the wild glutamine synthetase, the glutamine synthetase mutant disclosed by the invention has mutation at the nth site, wherein D, E, G, H, N, P, Q and V are formed or deleted after mutation, so that the glutamine synthetase by the mutation hasglufosinate-ammonium resistance. Thus, the glutamine synthetase mutant can be used for cultivating a new glufosinate-resistant plant variety.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a plant-derived glutamine synthetase mutant with glufosinate-ammonium resistance, nucleic acid molecules and applications. Background technique [0002] Glutamine synthetase (Glutamine synthetase, GS) is a key enzyme in plant nitrogen metabolism. It catalyzes the condensation of glutamic acid (Gln) and NH3 to form glutamine (Glu) in the glutamate synthetase cycle, and participates in plant nitrogen storage. Compound metabolism. According to the distribution and subcellular localization, the isoenzymes of higher plant GS (genus GSII) can be divided into two types: one is located in the cytoplasm and is called cytoplasmic GS (GS1), with a molecular weight of 38-40kDa; the other is located in the cytoplasm. The chloroplast (or plastid) is called plastid type GS (GS2), with a molecular weight of 44-45kDa. [0003] Glufosinate ammonium (glufosinate, glufosinate ammoniμM,...

Claims

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Application Information

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IPC IPC(8): C12N9/00C12N15/52C12N15/82A01H5/00A01H6/54A01H6/46A01H6/20
CPCC12N9/93C12Y603/01002C12N15/8277Y02A40/146
Inventor 邓龙群陈容张震候清江胥南飞
Owner SICHUAN GEVOTO BIOTECH CO LTD
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