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Plants having increased tolerance to herbicides

A herbicide and plant technology, applied in the direction of herbicides and algicides, plant growth regulators, botany equipment and methods, can solve problems that have not yet been described

Pending Publication Date: 2020-10-20
BASF SE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Thus, to date, the prior art has not described uracil-pyridine-tolerant plants containing the mutant PPO nucleic acid of the invention, which plants are tolerant / resistant to a wide range of uracil-pyridine options

Method used

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  • Plants having increased tolerance to herbicides
  • Plants having increased tolerance to herbicides
  • Plants having increased tolerance to herbicides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[2113] Example 1: Site-directed mutagenesis of PPO

[2114] All nucleic acid coding sequences and all single mutants and double mutants encoding herbicide-tolerant PPO polypeptides were synthesized and cloned by Geneart (Geneart AG, Regensburg, Germany). The rationally designed mutant is synthesized by Geneart. The random PPO gene library is synthesized by Geneart. By performing plasmid miniprep, the plasmid was isolated from E. coli TOP10 and confirmed by DNA sequencing.

Embodiment 2

[2115] Example 2: Expression and purification of recombinant wild-type and mutant PPO

[2116] (Taken from: Franck E. Dayan, Pankaj R. Daga, Stephen O. Duke, Ryan M. Lee, Patrick J. Tranel, Robert J. Doerksen. Biochemistry of glycine deletion in the α-8 helix of protoporphyrinogen oxidase Results and structural consequences (Biochemical and structural consequences of a glycinedeletion in the α-8 helix of protoporphyrinogen oxidase), Biochimica etBiophysica Acta 1804 (2010), 1548-56). The clone in pRSET vector was transformed into BL21(DE3)-pLysS E. coli strain. Place the cells in 100μg mL -1 Incubate in 250 mL LB with carbenicillin and shake overnight at 37°C. The culture was diluted in 1 L of antibiotic-containing LB and incubated at 37°C with shaking for 2 hours, induced with 1 mM IPTG and incubated at 25°C with shaking for another 5 hours. The cells were harvested by centrifugation at 1600×g, washed with 0.09% NaCl, and stored at -80°C. Use a French crusher at 140MPa in 50m...

Embodiment 3

[2117] Example 3: PPO enzyme assay (non-recombinant)

[2118] As previously mentioned, PPO protein (EC 1.3.3.4) was extracted from coleoptiles or seedlings (150 g fresh weight) of corn, nightshade, morning glory, and abutilon seedlings grown in the dark (Grossmann et al., 2010). Before harvest, the seedlings were allowed to turn green for 2 hours under light to achieve the highest specific enzyme activity in the thylakoid fraction at low chlorophyll concentrations. Obvious fluorescence quenching occurs at high chlorophyll concentrations, which limits the amount of green thylakoids that can be used in the experiment. Using a fresh weight to volume ratio of 1:4, the plant material was homogenized in a cold homogenization buffer with a Braun mixer. The homogenization buffer consists of tris(hydroxymethyl)aminomethane (Tris)-HCl (50mM; pH 7.3), sucrose (0.5M), magnesium chloride (1mM), ethylenediaminetetraacetic acid (EDTA) (1mM) and bovine serum white Egg white (2g L -1 )compositi...

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Abstract

The present invention refers to a method for controlling undesired vegetation at a plant cultivation site, the method comprising the steps of providing, at said site, a plant that comprises at least one nucleic acid comprising a nucleotide sequence encoding protoporphyrinogen oxidase (PPO) which is resistant or tolerant to a PPO-inhibiting herbicide by applying to said site an effective amount ofsaid herbicide. The invention further refers to plants comprising wild-type or mutated PPO enzymes, and methods of obtaining such plants.

Description

[0001] Field of invention [0002] The present invention generally relates to methods of imparting agricultural-level herbicide tolerance to plants. In particular, the present invention relates to plants having increased tolerance to PPO inhibiting herbicides. More specifically, the present invention relates to methods of mutagenesis and cross-breeding and transformation and to plants obtained by mutagenesis and cross-breeding and transformation, said plants having increased tolerance to PPO-inhibiting herbicides. [0003] Background of the invention [0004] Since the 1960s, herbicides that inhibit protoporphyrinogen oxidase (hereinafter referred to as protoporphyrinogen oxidase or PPO; EC: 1.3.3.4), a key enzyme in the biosynthesis of protoporphyrin IX, Agents have been used for selective weed control. PPO catalyzes the final common step in chlorophyll and heme biosynthesis, the oxidation of protoporphyrinogen IX to protoporphyrin IX. (Maringe et al., 1989. Biochem. 1.260:...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N43/54
CPCA01N43/54C12N9/001A01P13/00C12N15/8274C12Y103/03004
Inventor R·阿朋特T·塞萨尔S·特雷施P·A·鲍尔曼L·帕拉拉帕多M·维切尔L·苏伊拉尔M·约翰尼斯T·米茨纳J·M·保利克
Owner BASF SE
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