Compositions and methods for control of insect infestations in plants

An insect and plant technology, applied in the field of genetic control and modification of endogenous expression of coding sequences in cells or tissues of specific harmful organisms, can solve the problems of instability of antisense sequences, instability of antisense sequences, etc.

Active Publication Date: 2007-07-11
MONSANTO TECH LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

First, the antisense sequences expressed in transformed cells are unstable
Second, the instability of the antisense sequence expressed in the transformed cell subsequently creates difficulties in transporting that sequence to a host, cell type or biological system distant from the transgenic cell
It is believed to be impractical to provide dsRNA molecules in the diet of most invertebrate pest species or to inject compositions containing dsRNA into invertebrate pests

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0217] This example describes the identification of nucleotide sequences that are useful for controlling corn rootworm when provided in the corn rootworm diet in the form of double-stranded RNA molecules.

[0218] Corn rootworm cDNA libraries (LIB149, LIB150, LD33027, LIB3373) were constructed from whole larvae and from dissected midgut sections to obtain nucleotide sequence information (see, Andersen et al., submitted on July 24, 2002 10 / 205,189, dated U.S. Patent Application Serial No. 10 / 205,189, which is hereby expressly incorporated 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 to maximize the number of distinct EST sequences from Diabrotica species. Libraries LIB5444 and LIB5462 were constructed from mRNA libraries obtained from first instar (1 g) and third instar (2.9 g) Western corn rootworm larvae, respectively. The resulting insect...

Embodiment 2

[0321] This example describes significant pest suppression achieved by feeding an invertebrate pest a diet containing a double stranded RNA sequence obtained from the pest.

[0322] An artificial diet sufficient to rear corn rootworm larvae was prepared by using samples of double-stranded RNA sequences obtained from six different corn rootworm cDNA library sequences. Corn rootworm larvae were allowed to eat the diet for several days, and compared with corn rootworms allowed to eat only the control diet, mortality, morbidity and stunting were monitored. The nucleotide sequence used in the diet was obtained from the sequences shown in SEQ ID NO: 35, SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 52, SEQ ID NO: 7 and SEQ ID NO: 31 , each of which corresponds to the nucleotide sequence obtained from the corn rootworm cDNA library, and their deduced amino acid sequence translations correspond to the annotated 40kDa V-ATPase homologue, EF1α homologue, 26S proteosome subunit homologue, res...

Embodiment 3

[0336] This example describes nucleotide sequences for expression in plant cells, and the effect of providing such nucleotide sequences in a corn rootworm diet.

[0337]The CHD3 coding sequence obtained from a corn rootworm cDNA library was used to construct the nucleotide sequence encoding the stabilized double-stranded RNA. The cDNA sequence shown in SEQ ID NO: 171, encoding an ortholog or a portion of a CHD3 amino acid sequence ortholog, was used to construct primer pairs for thermal amplification reactions using the corn rootworm genome Template DNA. The primer pair set forth in SEQ ID NO:5 and SEQ ID NO:6 is capable of amplifying double-stranded genomic amplicons, one strand of which exhibits the sequence set forth in SEQ ID NO:7. Three nucleotide sequence fragments were generated from the nucleotide sequence shown in SEQ ID NO:7. Using the nucleotide sequence shown in SEQ ID NO: 7 as a template, in a thermal amplification reaction, with a pair of thermal amplification ...

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PUM

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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

[0001] Cross References to Related Applications [0002] This application claims the following U.S. provisional applications: 60 / 560,842 filed April 9, 2004; 60 / 565,632 filed April 27, 2004; 60 / 579,062 filed June 11, 2004; 60 / 603,421 dated August 20, 2004, 60 / 617,281 filed October 11, 2004, and 60 / _______ filed April 7, 2005, each of which is incorporated herein by reference in its entirety. field of invention [0003] In general, the present invention relates to the genetic control of pest infestation in plants as well as in and on animals. More specifically, the present invention relates to methods for modifying the endogenous expression of coding sequences in cells or tissues of specific pests. More specifically, the present invention uses recombinant DNA technology to post-transcriptionally repress or inhibit the expression of a target coding sequence in a pest cell by feeding the pest one or more genes transcribed from part or all of the target coding sequence. Invasion...

Claims

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Application Information

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IPC IPC(8): C12N15/09C12N15/82C12N15/31A01H5/00
Inventor J·A·鲍姆L·A·吉尔伯特森D·K·科瓦利克T·J·拉罗萨M·卢T·R·I·蒙伊克瓦J·K·罗伯茨W·吴B·张
Owner MONSANTO TECH LLC
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