Compositions and methods for control of insect infestations in plants

a technology for insects and plants, applied in the field of genetic control of pest infestations in plants, can solve the problems of slow metabolism, commercial crops are often targets, and use chemical pesticidal agents, and achieve the effect of limiting or eliminating invertebra

Inactive Publication Date: 2006-01-26
MONSANTO TECH LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0013] The present invention, in one embodiment, comprises a method of inhibiting expression of a target gene in an invertebrate pest. Specifically, the present invention comprises a method of modulating or inhibiting expression of one or more target genes in an invertebrate pest, in particular, in lygus bugs (Lygus hesperus Knight) and the like, that cause cessation of feeding, growth, development, reproduction and infectivity and eventually result in the death of the insect. The method comprises introduction of partial or fully, stabilized double-stranded RNA (dsRNA) or its modified forms such as small interfering RNA (siRNA) sequences, into the cells or into the extracellular environment, such as the midgut, within...

Problems solved by technology

Commercial crops are often the targets of insect attack.
However, there are several disadvantages to using chemical pesticidal agents.
However, because of the lack of selectivity, the chemical pesticidal agents exert their effects on non-target fauna as well, often effectively sterilizing a field for a period of time over which the pesticidal agents have been applied.
Chemical pesticidal agents persist in the environment and generally are slow to be metabolized, if at all.
Accumulations of these chemical pesticidal agents results in the development of resistance to the agents and in species higher up the evolutionary ladder, act as mutagens and/or carcinogens often causing irreversible and deleterious genetic modifications.
Antisense technology has been difficult to employ in many systems for three principle reasons.
First, the antisense sequence expressed in the transformed cell is unstable.
Second, the instability of the antisense sequence expressed in the transformed cell concomitantly creates difficulty in delivery of the sequence to a host, cell type, or biological system remote from the transgenic cell.
Third, the difficulties encountered with instability and delivery of the antisense sequence create difficulties in ...

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0194] This example illustrates the identification of nucleotide sequences that, when provided in the double stranded RNA form in the diet of a lygus species insect pest, are useful for controlling a lygus species insect pest.

[0195]Lygus species, and in particular Lygus hesperus, are typical of a class of pests that infest crop plants and ornamentals and globally cause severe commercially significant levels of damage on an annual basis. This class of pests effect their damage by piercing the defenses of a plant, plant tissue, or plant cell and subsequently extract nutritional value from the plants. Nutritional value is captured by the pest using this means of attack by one of two or three modes of action. One means is to puncture or penetrate into individual cells and suck out the juices of those cells. Another means is to penetrate into the xylem or phloem vesicles of the plant or plant tissues and allow the turgor pressure within the plant to extrude nutritionally rich xylem or p...

example 2

[0203] This example illustrates the identification of nucleotide sequences that, when provided in the form of double stranded RNA molecules in the diet of a corn rootworm, are useful for controlling corn rootworms.

[0204] Corn rootworm cDNA libraries (LIB149, LIB 150, LIB3027, LIB3373) were constructed from whole larvae and from dissected midgut sections, and nucleotide sequence information was obtained (see Andersen et al., U.S. patent application Ser. No. 10 / 205,189 filed Jul. 24, 2002, incorporated herein specifically by reference in its entirety). In addition, cDNA libraries were constructed from whole larvae at different developmental stages and at different times within each developmental stage in order to maximize the number of different EST sequences from the Diabrotica species. Libraries LIB5444 and LIB5462 were constructed respectively from mRNA pools obtained from first (1 gram) and third (2.9 grams) instar Western Corn Rootworm larvae. Harvested insects were rapidly froz...

example 3

[0288] This example illustrates significant pest inhibition obtained by feeding to an invertebrate pest a diet containing double stranded RNA sequences derived from that pest.

[0289] Artificial diet sufficient for rearing corn rootworm larvae was prepared by applying samples of double stranded RNA sequences derived from six different corn rootworm cDNA library sequences. Corn rootworm larvae were allowed to feed on the diet for several days and mortality, morbidity and stunting monitored in comparison to rootworms allowed to feed only on control diet. The nucleotide sequences that were used in the diet were derived from sequences as set forth in SEQ ID NO:46, SEQ ID NO:50, SEQ ID NO:58, SEQ ID NO:63, SEQ ID NO:18, and SEQ ID NO:42, each corresponding to nucleotide sequences derived from a corn rootworm cDNA library, the deduced amino acid sequence translation of which corresponds respectively to proteins annotated to a 40 kDa V-ATP-ase homolog, an EF1α homolog, a 26S proteasome subu...

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Abstract

The present invention is directed to controlling pest infestation by inhibiting one or more biological functions in an invertebrate pest. The invention discloses methods and compositions for use in controlling pest infestation by feeding one or more different recombinant double stranded RNA molecules to the pest in order to achieve a reduction in pest infestation through suppression of gene expression. The invention is also directed to methods for making transgenic plants that express the double stranded RNA molecules, and to particular combinations of transgenic pesticidal agents for use in protecting plants from pest infestation.

Description

CROSS REFERENCE TO RELATED APPLICATIONS [0001] This application claims priority to U.S. Provisional Application Nos. 60 / 560,842, filed Apr. 9, 2004, 60 / 565,632, filed Apr. 27, 2004, 60 / 579,062, filed Jun. 11, 2004, 60 / 603,421, filed Aug. 20, 2004, 60 / 617,261, filed Oct. 11, 2004, and 60 / ______, filed on Apr. 7, 2005.FIELD OF THE INVENTION [0002] The present invention relates generally to genetic control of pest infestations in plants and in and on animals. More specifically, the present invention relates to the methods for modifying endogenous expression of coding sequences in the cell or tissue of a particular pest. More specifically, the present invention utilizes recombinant DNA technologies to post-transcriptionally repress or inhibit expression of a target coding sequence in the cell of a pest, by feeding to the pest one or more double stranded or small interfering ribonucleic acid (RNA) molecules transcribed from all or a portion of a target coding sequence, thereby controllin...

Claims

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Application Information

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IPC IPC(8): A01H1/00C12N5/04C07H21/04C12N15/82A01N63/02A23L19/00
CPCC12N15/8286C12N15/8279C12N15/8285A01N63/50A01N63/60Y02A40/146A01N63/23C12N15/8275
Inventor BAUM, JAMESGILBERTSON, LARRYKOVALIC, DAVIDLAROSA, THOMASLU, MAOLONGMUNYIKWA, TICHAFAROBERTS, JAMESWU, WEIZHANG, BEI
Owner MONSANTO TECH LLC
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