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Glutamine synthetase mutant and application of glutamine synthetase mutant in culturing glufosinate-ammonium-resistant plant variety

A glutamine and synthetase technology, applied in the field of genetic engineering, can solve the problems of affecting plant growth and development, glufosinate-ammonium tolerance is not enough for commercial application, interfering with plant nitrogen metabolism, etc., to achieve the effect of satisfying growth and development

Pending Publication Date: 2021-11-05
SICHUAN GEVOTO BIOTECH CO LTD
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

However, before glufosinate-ammonium contacts glutamine synthetase, it is difficult for glufosinate-ammonium acetylase to completely inactivate glufosinate-ammonium. Inhibits the activity of glutamine synthetase on the cell membrane, thereby interfering with plant nitrogen metabolism
Therefore, when glufosinate-ammonium is applied to crops with bar gene and pat gene, it will interfere with the nitrogen metabolism of plants to varying degrees, and at the same time affect the normal growth and development of plants.
Although the sensitivity of transgenic plants to glufosinate can be reduced to some extent by overexpressing wild-type glutamine synthetase in plants, their tolerance to glufosinate is far from enough for commercial application

Method used

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  • Glutamine synthetase mutant and application of glutamine synthetase mutant in culturing glufosinate-ammonium-resistant plant variety
  • Glutamine synthetase mutant and application of glutamine synthetase mutant in culturing glufosinate-ammonium-resistant plant variety
  • Glutamine synthetase mutant and application of glutamine synthetase mutant in culturing glufosinate-ammonium-resistant plant variety

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0099] The rice (Oryza sativa) glutamine synthetase (GS1) mutant provided in this embodiment is composed of the wild-type rice glutamine synthetase itself (named OWT, the amino acid sequence is shown in SEQ ID NO.1, and the encoding nucleus The nucleotide sequence is that the 69th amino acid residue E of SEQ ID NO.6) is mutated into A, C, D, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, Y or deletion, the obtained rice GS1 mutants were named OE69A, OE69C, OE69D, OE69F, OE69G, OE69H, OE69I, OE69K, OE69L, OE69M, OE69N, OE69P, OE69Q, OE69R, OE69S, OE69T, OE69V, OE69Y, and OE69X.

[0100] Comparison of amino acid sequences of rice GS1 mutants OE69A, OE69C, OE69D, OE69F, OE69G, OE69H, OE69I, OE69K, OE69L, OE69M, OE69N, OE69P, OE69Q, OE69R, OE69S, OE69T, OE69V, OE69Y and OE69X with wild-type rice GS1 For example figure 1 As shown in the figure: the position indicated by the arrow is the mutation site.

[0101] In this example, the coding sequence of each rice GS1 mutant is at the po...

Embodiment 2

[0105] The soybean (Glycine max) GS1 mutant provided in this example is composed of the wild-type soybean GS1 itself (named GWT, the amino acid sequence is shown in SEQ ID NO.3, and the encoding nucleotide sequence is SEQ ID NO.8). The 69th position (corresponding to the 69th position of the reference sequence (SEQ ID NO.1)) is obtained by mutating the amino acid residue E to A, G, L, M, N, Q, S, T, V or deleting it. The obtained soybean GS1 mutants were named GE69A, GE69G, GE69L, GE69M, GE69N, GE69Q, GE69S, GE69T, GE69V and GE69X, respectively.

[0106] The amino acid sequence alignment of soybean GS1 mutants GE69A, GE69G, GE69L, GE69M, GE69N, GE69Q, GE69S, GE69T, GE69V and GE69X and wild-type soybean GS1 GWT is as follows: figure 2 As shown in the figure: the position indicated by the arrow is the mutation site.

[0107] The coding sequences of soybean GS1 mutants GE69A, GE69G, GE69L, GE69M, GE69N, GE69Q, GE69S, GE69T, GE69V and GE69X provided in this example correspond to...

Embodiment 3

[0112] The maize (Zea mays) GS1 mutant provided in this example is composed of the wild-type maize GS1 itself (named ZWT, the amino acid sequence is shown in SEQ ID NO.2, and the encoding nucleotide sequence is SEQ ID NO.7). The 69th position (corresponding to the 69th position of the reference sequence (SEQ ID NO.1)) is mutated from amino acid residue E to A, C, D, F, G, H, I, K, L, M, N, P , Q, R, S, T, V, Y or deleted. The maize GS1 mutants obtained were named ZE69A, ZE69C, ZE69D, ZE69F, ZE69G, ZE69H, ZE69I, ZE69K, ZE69L, ZE69M, ZE69N, ZE69P, ZE69Q, ZE69R, ZE69S, ZE69T, ZE69V, ZE69Y and ZE69X.

[0113] Amino acid sequence comparison of maize GS1 mutants ZE69A, ZE69C, ZE69D, ZE69F, ZE69G, ZE69H, ZE69I, ZE69K, ZE69L, ZE69M, ZE69N, ZE69P, ZE69Q, ZE69R, ZE69S, ZE69T, ZE69V, ZE69Y and ZE69X and wild type maize GS1ZW For example image 3 As shown in the figure: the position indicated by the arrow is the mutation site.

[0114] In this example, the coding sequence of each maize...

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Abstract

The invention discloses a glutamine synthetase mutant and application of the glutamine synthetase mutant in cultivation of a glufosinate-ammonium-resistant plant variety. The inventor discovers that the glutamine synthetase mutant can be obtained by mutation at the nth site of wild glutamine synthetase, the mutant is A, G, M, N, Q, S, T and V or deleted after mutation, and the mutation can endow the glutamine synthetase with glufosinate-ammonium resistance suitable for commercial application. The glutamine synthetase mutant has the application potential of constructing an expression vector for transforming plants and cultivating glufosinate-ammonium-resistant crops.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a glutamine synthetase mutant and its application in cultivating glufosinate-resistant plant varieties. Background technique [0002] Glufosinate ammonium (glufosinate ammonium, trade name Basta) is a glutamine synthetase (GS1) inhibitor developed by Bayer. Its active ingredient is phosphinothricin (abbreviated as PPT), and its chemical name is (RS)-2-amino-4 - Ammonium (hydroxymethylphosphinyl)butyrate. By inhibiting the activity of glutamine synthetase (Glutamine synthetase, GS), the synthesis of glutamine in plants is blocked, and then the nitrogen metabolism in plants is disordered, the synthesis of proteins and nucleotides and other substances is reduced, and photosynthesis is blocked. Chlorophyll synthesis decreased. At the same time, the content of ammonium ions in the cells increases, causing the cell membranes to be damaged, the chloroplasts to disintegrate...

Claims

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Application Information

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IPC IPC(8): C12N9/00C12N15/52C12N15/82A01H5/00A01H6/46A01H6/82A01H6/54A01H6/20A01H6/60
CPCC12N9/93C12N15/8277C12Y603/01002
Inventor 陈容侯青江邓龙群张震胥南飞
Owner SICHUAN GEVOTO BIOTECH CO LTD
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