Cry Toxin Gene Variants for Lepidopteran Pest Control

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Solution Overview

Problem

There is a continuous need for new forms of pesticidal toxins to control insect pests effectively in agriculture, as existing methods may not fully address the devastation caused by insects and the need for improved yield protection.

Innovation Solution

The development of novel nucleic acid molecules encoding pesticidal proteins, which can be used to transform bacteria, plants, and seeds, providing them with pesticidal activity against lepidopteran, hemipteran, coleopteran, nematode, or dipteran pests, and methods for detecting these proteins and nucleic acids.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If existing pesticidal toxins are used to control insect pests, then some pest control is achieved, but the effectiveness is insufficient to fully address the devastation caused by insects

Engineering Contradiction:
Improvepest control effectivenessVSAvoidagricultural yield protection
Core Design Contradiction:
ReliabilityVSProductivity

Solution Approach 1:

The patent modifies existing Cry toxin genes through site-directed mutagenesis to create variant toxins with improved insecticidal activity. Specific amino acid substitutions are introduced to enhance binding affinity to insect midgut receptors, thereby improving the reliability of pest control while maintaining productivity benefits

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent creates composite pesticidal compositions by combining multiple Cry toxin variants with different insecticidal spectra. This allows simultaneous control of multiple pest species (Lepidoptera, Diptera, Coleoptera) that cannot be effectively controlled by single toxins, thereby improving overall pest control effectiveness and agricultural yield protection

Inventive Principle:
Principle #40Composite materials

2Reliability

If new pesticidal toxin forms are developed, then pest control effectiveness is improved, but the complexity of discovering and characterizing new toxins increases

Engineering Contradiction:
Improvepest control effectivenessVSAvoidtoxin discovery and characterization process
Core Design Contradiction:
ReliabilityVSDevice complexity

Solution Approach 1:

The patent performs preliminary site-directed mutagenesis on well-characterized Cry toxin genes to generate variant toxins with predicted improved activity. This preliminary molecular modification approach allows systematic exploration of structure-activity relationships without the need for de novo toxin discovery, thereby reducing the complexity of the development process while improving pest control effectiveness

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The patent creates multiple copies of modified toxin genes and expresses them in different host systems (bacteria, plants) to validate and scale up the pesticidal activity. This copying and validation approach simplifies the characterization process by using established molecular biology techniques rather than requiring entirely new toxin isolation and characterization methodologies

Inventive Principle:
Principle #26Copying

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

The novel pesticidal proteins confer enhanced pest resistance or tolerance to organisms, enabling effective control of target pest populations and improving agricultural yield by reducing pest infestation and damage.

Implementation Method 1

This toxin binds to apical brush border receptors in the midgut of the target larvae and inserts into the apical membrane creating ion channels or pores, resulting in larval death

Methodology Applied
Scientific EffectIon channel formation:

Implementation Method 2

The ingested protoxin is hydrolyzed by proteases in the insect digestive tract to an active toxic molecule

Methodology Applied
Scientific EffectProteolytic hydrolysis: Hydrolysis

Data Source

PatentUS10221431B2AXMI422 toxin gene and methods for its use
Publication Date: 2019.03.05 BASF AGRICULTURAL SOLUTIONS US LLC

AI summary

Compositions and methods for conferring pesticidal activity to bacteria, plants, plant cells, tissues and seeds are provided. Compositions comprising a coding sequence for a toxin polypeptide are provided. The coding sequences can be used in DNA constructs or expression cassettes for transformation and expression in plants and bacteria. Compositions also comprise transformed bacteria, plants, plant cells, tissues, and seeds. In particular, isolated toxin nucleic acid molecules are provided. Additionally, amino acid sequences corresponding to the polynucleotides are encompassed, and antibodies specifically binding to those amino acid sequences. In particular, the present invention provides for isolated nucleic acid molecules comprising nucleotide sequences encoding the amino acid sequence shown in SEQ ID NO:2-3, or the nucleotide sequence set forth in SEQ ID NO: 1, as well as variants and fragments thereof.