Grg33, grg35, grg36, grg37, grg38, grg39, and grg50: novel epsp synthase genes conferring herbicide resistance

A herbicide and resistance technology, applied in the direction of plant genetic improvement, botany equipment and methods, biochemical equipment and methods, etc.

Inactive Publication Date: 2009-07-08
ATHENIX
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the EPSP synthases of certain bacteria are highly tolerant to glyphosate

Method used

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  • Grg33, grg35, grg36, grg37, grg38, grg39, and grg50: novel epsp synthase genes conferring herbicide resistance
  • Grg33, grg35, grg36, grg37, grg38, grg39, and grg50: novel epsp synthase genes conferring herbicide resistance
  • Grg33, grg35, grg36, grg37, grg38, grg39, and grg50: novel epsp synthase genes conferring herbicide resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0091] Example 1. Isolation of glyphosate-resistant EPSP synthase

[0092] Strains capable of growing in the presence of glyphosate were isolated by plating soil samples on HEPES mineral salt medium (HMSM) containing glyphosate as the sole phosphorus source. Because HMSM does not contain aromatic amino acids, strains must be resistant to glyphosate in order to grow on this medium.

[0093] 2 grams of soil were resuspended in approximately 10 ml of water, vortexed for 15 seconds, and allowed to settle for 15 minutes. A 10 μl loop of this suspension was added to 3 ml HMSM (pH 7.0) supplemented with 10 mM glyphosate. HMSM contains (per liter): 10g glucose, 2gNH 4 SO 4 , 9.53g HEPES, 1.0ml 0.8M MgSO 4 , 1.0ml 0.1M CaCl 2 , 1.0ml trace element solution (in 100ml of 1000x solution: 0.1g FeSO 4 ·7H 2 O, 0.5mg CuSO 4 ·5H 2 O, 1.0 mg H 3 BO 3 , 1.0mg MnSO 4 ·5H 2 O, 7.0mg ZnSO 4 ·7H 2 O, 1.0mg MoO 3 , 4.0 g KCl). The culture was grown for 4 days at 28°C in a shaking ...

Embodiment 2

[0096] Example 2. Isolation of glyphosate-resistant EPSP synthases grg37 and grg39

[0097] Strains capable of growing in the presence of glyphosate were isolated by plating soil samples on various growth media containing glyphosate. Certain strains were isolated on mineral salt media supplemented with glyphosate. Other strains were isolated in rich media in the presence of glyphosate and subsequently tested on mineral salt media supplemented with glyphosate. Because mineral salt medium does not contain aromatic amino acids, strains must be resistant to glyphosate in order to grow on this medium.

[0098] Strains ATX21800 and ATX21804 were isolated by incubation under enriched conditions and supplemented with glyphosate. These strains were then tested for their ability to grow in the presence of glyphosate and without aromatic amino acids. Strain ATX21804 was isolated from soil (0.01 g) air-dried for 2 days and plated on nutrient broth agar supplemented with 100 mM glypho...

Embodiment 3

[0102] Example 3. Isolation of glyphosate-resistant EPSP synthases grg38 and grg50

[0103] Strains capable of growing in the presence of glyphosate were isolated by plating soil samples on various growth media containing glyphosate. Certain strains were isolated on mineral salt media supplemented with glyphosate. Other strains were isolated in rich media in the presence of glyphosate and subsequently tested on mineral salt media supplemented with glyphosate. Because mineral salt medium does not contain aromatic amino acids, strains must be resistant to glyphosate in order to grow on this medium.

[0104] Strain ATX20103 was isolated by resuspending approximately 2 grams of soil in approximately 10 ml of water, vortexing for 15 seconds, and allowing it to settle for 15 minutes. A 10 μl loop of this suspension was added to 3 ml Tris MSM (TMSM) (pH 7.0) supplemented with 10 mM glyphosate. TMSM contains (per liter): 10g glucose, 2g NH 4 SO 4 , 12.12g Tris, 1.0ml 0.8M MgSO ...

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Abstract

Compositions and methods for conferring herbicide resistance to bacteria, plants, plant cells, tissues and seeds are provided. Compositions include nucleic acid molecules encoding herbicide resistance or tolerance polypeptides, vectors comprising those nucleic acid molecules, and host cells comprising the vectors. The nucleotide sequences of the invention can be used in DNA constructs or expression cassettes for transformation and expression in organisms, including microorganisms and plants. Compositions also comprise transformed bacteria, plants, plant cells, tissues, and seeds. In particular, the present invention provides for isolated nucleic acid molecules comprising the nucleotide sequence set forth in SEQ ID NO: 1, 3, 4, 6, 1, 9, 10, 12, 13, 15, 17, 18, 20, 21, or 23, a nucleotide sequence encoding the amino acid sequence shown in SEQ ID NO:2, 5, 8, 11, 14, 16, 19, or 22, the herbicide resistance nucleotide sequence deposited in a bacterial host as Accession Nos. NRRL B-30932, B-30933, B-30934, B-30945, B-30946, B-30947, or B-30948, as well as variants and fragments thereof.

Description

field of invention [0001] The present invention provides a novel gene encoding 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase, which confers herbicide resistance. These genes can be used in plant biology, crop breeding and plant cell culture. Background of the invention [0002] N-phosphonomethylglycine, commonly known as glyphosate, is an important agronomic compound. Glyphosate inhibits the enzyme that converts phosphoenolpyruvate (PEP) and 3-phosphoshikimate to 5-enolpyruvyl-3-phosphoshikimate. Inhibition of this enzyme (5-enolpyruvylshikimate-3-phosphate synthase; referred to herein as "EPSP synthase") inhibits aromatase by shutting down the shikimate pathway. Biosynthesis of family amino acids kills plant cells. [0003] Because glyphosate herbicides inhibit the biosynthesis of aromatic amino acids, they not only kill plant cells but are also toxic to bacterial cells. Glyphosate inhibits the EPSP synthase of many bacteria and is therefore toxic to these bacteria...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H1/00C12Q1/68
Inventor C·L·皮特斯B·范德伯格B·卡尔D·J·汤姆索
Owner ATHENIX
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