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Compositions and methods for controlling diabrotica

a technology of diabrotica and diabrotica, which is applied in the field of methods for controlling invertebrate pest infestations, can solve the problems of chemical insecticide use and potential accumulation in the food chain, and achieve the effect of preventing infestation and low nucleotide diversity

Inactive Publication Date: 2016-08-11
MONSANTO TECH LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patent is about controlling pests, specifically Diabrotica species, which are important economic pests. The invention relates to using recombinant DNA techniques to create plants that are resistant to infestation by these pests. The patent describes methods for making plants that express specific polynucleotides or apply topical agents containing polynucleotides to prevent infestation. The invention also includes identifying target genes in the pest that can be targeted for RNA interference to control infestation. Overall, the patent provides techniques for creating plants that are more resistant to Diabrotica species infestation.

Problems solved by technology

Commercial crops are often the targets of attack by invertebrate pests such as insects.
However, there are several disadvantages to using chemical insecticides.
For example, chemical insecticides are generally not selective, and applications of chemical insecticides intended to control insect pests in crop plants can exert their effects on non-target insects and other invertebrates as well.
Chemical insecticides often persist in the environment and can be slow to degrade, thus potentially accumulating in the food chain.

Method used

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  • Compositions and methods for controlling diabrotica

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0271]This example illustrates non-limiting embodiments of sequences useful as target genes for controlling Diabrotica species and for making compositions and plants of this invention, and describes identification of such DNA sequences from Diabrotica virgifera virgifera. It is recognized that analogous sequences can be obtained from any other Diabrotica species referred to hereinabove.

[0272]cDNA libraries were generated from mid-guts of Diabrotica virgifera virgifera (Western corn rootworm, WCR) third instar larvae reared on corn plants, as follows. Disinfected WCR eggs were suspended in a 0.1% (w / v) agar solution and dispensed into petri dishes containing 2% (w / v) agar and filter paper. The WCR eggs were incubated first at 20 degrees Celsius and 60% relative humidity (“RH”) for 3 days and then at 25 degrees Celsius and 60% RH for 10 days. After 13 days of incubation, the eggs were washed from the dishes into sweater boxes containing soil (2 parts Metro-Mix 200: 1 part Redi-Earth; ...

example 2

[0273]This example illustrates a method for non-random selection of target genes. More specifically, this example illustrates a method of selecting a subset of target genes that are present in single- or low-copy-number (i. e., non-repetitive and non-redundant) in a particular genome.

[0274]In general the method includes the step of identifying single- or low-copy-number genes in the chosen genome, or alternatively to identify single- or low-copy-number genes in an orthologous database from related organisms to predict which genes will be single / low copy in the chosen organism. Low-copy genes, and in particular single-copy genes, are unlikely to have their function recapitulated by a paralogue, and are selected as preferred targets for RNAi-mediated silencing. In other words, such genes are likely to represent essential functions, making these genes preferred targets for RNAi-mediated silencing.

[0275]Embodiments of the method include a further step of estimating nucleotide diversity ...

example 3

[0282]The polynucleotides of this invention are generally designed to modulate expression by inducing regulation or suppression of a Diabrotica species target gene and are designed to have a nucleotide sequence essentially identical or essentially complementary to the nucleotide sequence of a Diabrotica species target gene (e. g., SEQ ID NOs:1-450) or to the sequence of RNA transcribed from a Diabrotica species target gene, which can be coding sequence or non-coding sequence. These effective polynucleotide molecules that modulate expression are referred to herein as a “trigger”, or “triggers”. This example describes non-limiting techniques useful in the design and selection of polynucleotides as “triggers” to modulate expression of a Diabrotica species target gene.

[0283]Selection of Polynucleotide Triggers by “Tiling”

[0284]Polynucleotides of use in the invention need not be of the full length of a target gene, and in many embodiments are of much shorter length in comparison to the t...

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Abstract

The present invention provides methods for controlling invertebrate pest infestations, for instance in plants, and related compositions and polynucleotides useful in such methods. More specifically, the present invention provides polynucleotides and methods of use thereof for modifying the expression of genes in an invertebrate pest, for instance through RNA interference.

Description

CROSS REFERENCE TO RELATED APPLICATIONS[0001]This application claims the benefit of U.S. Provisional Application No. 61 / 782,884, filed Mar. 14, 2013, herein incorporated by reference in its entirety.INCORPORATION OF SEQUENCE LISTINGS[0002]The sequence listing that is contained in the file named “MONS355US.txt”, which is 699 kilobytes (as measured in Microsoft Windows®) and was created on Mar. 11, 2014, is filed herewith by electronic submission and is incorporated by reference herein.FIELD OF THE INVENTION[0003]This invention discloses methods for controlling invertebrate pest infestations, particularly in plants, and compositions and polynucleotides useful in such methods. More specifically, this invention is related to polynucleotides and methods of use thereof for modifying the expression of genes in an invertebrate pest, particularly through RNA interference. Particular pest species of interest include Diabrotica species, especially those that infest crop plants.BACKGROUND OF TH...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N15/82C12N15/113A01N57/16
CPCC12N15/8286C12N15/113A01N57/16C12N2310/14Y02A40/146
Inventor BAUM, JAMES A.BOLOGNESI, RENATAFLAGEL, LEX EVANSEGERS, GERRIT CORNELIS
Owner MONSANTO TECH LLC
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